Before scientist learned how to make a synthetic growth hormone, removing it painstakingly in small amounts from the pituitary glands of human cadavers. a. scientists ...
This article is about Frequently asked questions on electrical safety. Now that the new Part P Regulations have come in, what should I remember about
TY - JOUR. T1 - Saccharomyces cerevisiae proteins involved in hybrid DNA formation in vitro. AU - Heyer, W. D.. AU - Johnson, A. W.. AU - Norris, D. N.. AU - Tishkoff, D.. AU - Kolodner, R. D.. PY - 1991. Y1 - 1991. N2 - RecA-like activities that can form hybrid DNA in vitro have been identified in a wide variety of organisms. We have previously described the strand exchange protein 1 (SEP1) from the yeast Saccharomyces cerevisiae that can form hybrid DNA in vitro. Purified as an Mr 132 000 polypeptide, recent molecular and immunological studies have now shown that the native form is an Mr 175 000 polypeptide containing strand exchange activity. The gene encoding SEP1 has been cloned and sequenced. The primary sequence failed to reveal any significant sequence homology to other sequences in data base searches. In vivo SEP1 was found to be essential for normal meiosis as cells containing a homozygous insertion mutation in the SEP1 gene failed to sporulate. In order to identify additional factors ...
TY - JOUR. T1 - A regulated MET3-GLC7 gene fusion provides evidence of a mitotic role for Saccharomyces cerevisiae protein phosphatase 1. AU - Black, S. AU - Andrews, P D. AU - Sneddon, A A. AU - Stark, M J. PY - 1995. Y1 - 1995. N2 - Saccharomyces cerevisiae possesses a single essential gene (GLC7) encoding protein phosphatase 1 (PP1). Elevated expression of this gene from the GAL1 promoter is highly detrimental to the cell, causing a growth defect and aberrant bud morphology, which leads to cells exhibiting long, extended buds. By comparison, expression of GLC7 from the weaker MET3 promoter was without significant effect on either growth or morphology. However, repression of GLC7 expression from the MET3 promoter in cells where the MET3-GLC7 fusion was the sole source of PP1 resulted in a mitotic delay. Such cultures showed a massive decrease in the rate of proliferation in conjunction with a significant increase in the proportion of large, budded cells. 46-diamidino-2-phenylindole ...
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I reveal that Saccharomyces cerevisiae Rtt109p promotes genome stability and resistance to DNA-damaging agents, and that it does this by functionally cooperating with the histone chaperone Asf1p to maintain normal chromatin structure. Furthermore, I show that, as for Asf1p, Rtt109p is required for histone H3 acetylation on lysine 56 (K56) in vivo. Moreover I show that Rtt109p directly catalyzes this modification in vitro in a manner that is stimulated by Asf1p. These data establish Rtt109p as a member of a new class of histone acetyltransferases and show that its actions are critical fro cell survival in the presence of DNA damage during S phase. In the second part of this thesis, I reveal that cells deleted for Saccharomyces cerevisiae ESC2 exhibit synthetic sickness when combined with deletions of many genes involved in maintaining genomic stability. Moreover, I show that esc2Δ mutant cells exhibit increased recombination frequency and increased relocalisation of recombination repair protein ...
New Sequences ============= S82971 S82971 1775bp DNA PLN 10-FEB-1997 PEX13=PAS20 [Saccharomyces cerevisiae, Genomic, 1775 nt]. PEX13; Pex13p. SCRGA1 X90950 4305bp DNA PLN 07-FEB-1997 S.cerevisiae rga1 (dbm1) gene. DBM1; pheromone response; RGA1 gene; RGA1 (DBM1); Rga1p (Dbm1p). SCU17262 U17262 3051bp DNA PLN 11-FEB-1997 Saccharomyces cerevisiae Pip1p (PIP1) gene, complete cds. PIP1; Pip1p. SCU17263 U17263 2251bp DNA PLN 11-FEB-1997 Saccharomyces cerevisiae Pip2p (PIP2) gene, complete cds. PIP2; Pip2p. SCU17264 U17264 1842bp DNA PLN 11-FEB-1997 Saccharomyces cerevisiae Pip3p (PIP3) gene, complete cds. PIP3; Pip3p. SCU85960 U85960 1720bp DNA PLN 11-FEB-1997 Saccharomyces cerevisiae RNA polymerase II-specific TBP associated factor Taf40p (TAF40) gene, complete cds. TAF40; RNA polymerase II specific TBP associated; factor. SCU86641 U86641 1657bp DNA PLN 08-FEB-1997 Saccharomyces cerevisiae Rim9p (RIM9) gene, complete cds. RIM9; Rim9p. =========== Updated Features/Annotations ============= YSCDYS1 ...
TY - JOUR. T1 - Regulation and intracellular localization of Saccharomyces cerevisiae strand exchange protein 1 (Sep1/Xrn1/Kem1), a multifunctional exonuclease. AU - Heyer, Wolf Dietrich. AU - Johnson, Arlen W.. AU - Reinhart, Ursula. AU - Kolodner, Richard D.. PY - 1995/5. Y1 - 1995/5. N2 - The Saccharomyces cerevisiae strand exchange protein 1 (Sep1; also referred to as Xrn1, Kem1, Rar5, or Stpβ) catalyzes the formation of hybrid DNA from model substrates in vitro. The protein is also a 5-to-3 exonuclease active on DNA and RNA. Multiple roles for the in vivo function of Sep1, ranging from DNA recombination and cytoskeleton to RNA turnover, have been proposed. We show that Sep1 is an abundant protein in vegetative S. cerevisiae cells, present at about 80,000 molecules per diploid cell. Protein levels were not changed during the cell cycle or in response to DNA-damaging agents but increased twofold during meiosis. Cell fractionation and indirect immunofluorescence studies indicated that , 90% ...
TY - JOUR. T1 - Isolation and characterization of the RAD2 gene of Saccharomyces cerevisiae. AU - Higgins, David R.. AU - Prakash, Louise. AU - Reynolds, Paul. AU - Prakash, Satya. PY - 1984/10. Y1 - 1984/10. N2 - We have cloned the RAD2 gene of Saccharomyces cerevisiae and used it to determine the size and direction of its transcript and to make rad2 deletion mutants. The RAD2 gene encodes a 3.3-kb transcript and the direction of transcription is leftwards, from EcoRI towards BglII. Deletions of the RAD2 gene have no effect on viability of vegetative cells or spores, or on sporulation.. AB - We have cloned the RAD2 gene of Saccharomyces cerevisiae and used it to determine the size and direction of its transcript and to make rad2 deletion mutants. The RAD2 gene encodes a 3.3-kb transcript and the direction of transcription is leftwards, from EcoRI towards BglII. Deletions of the RAD2 gene have no effect on viability of vegetative cells or spores, or on sporulation.. KW - DNA repair. KW - ...
TY - JOUR. T1 - Molecular cloning and characterization of the RAD1 gene of Saccharomyces cerevisiae. AU - Higgins, David R.. AU - Prakash, Satya. AU - Reynolds, Paul. AU - Prakash, Louise. PY - 1983. Y1 - 1983. N2 - We have cloned the RAD1 gene of Saccharomyces cerevisiae and physically mapped it to a 4.0-kb DNA fragment from chromosome XVI. The RAD1 gene determines a transcript of 3.1 kb, and the direction of transcription was found to be leftwards, from EcoRI towards BglII (Fig. 1). Deletions of the RAD1 gene were made and were found to have no effect on viability of vegetative cells or spores, or on sporulation.. AB - We have cloned the RAD1 gene of Saccharomyces cerevisiae and physically mapped it to a 4.0-kb DNA fragment from chromosome XVI. The RAD1 gene determines a transcript of 3.1 kb, and the direction of transcription was found to be leftwards, from EcoRI towards BglII (Fig. 1). Deletions of the RAD1 gene were made and were found to have no effect on viability of vegetative cells or ...
Potential DNA replication accessory factors from the yeast Saccharomyces cerevisiae have previously been identified by their ability to bind to DNA polymerase alpha protein affinity matrices (J. Miles and T. Formosa, Proc. Natl. Acad. Sci. USA 89:1276-1280, 1992). We have now used genetic methods to characterize the gene encoding one of these DNA polymerase alpha-binding proteins (POB1) to determine whether it plays a role in DNA replication in vivo. We find that yeast cells lacking POB1 are viable but display a constellation of phenotypes indicating defective DNA metabolism. Populations of cells lacking POB1 accumulate abnormally high numbers of enlarged large-budded cells with a single nucleus at the neck of the bud. The average DNA content in a population of cells lacking POB1 is shifted toward the G2 value. These two phenotypes indicate that while the bulk of DNA replication is completed without POB1, mitosis is delayed. Deleting POB1 also causes elevated levels of both chromosome loss and ...
In the study, 300 male day-old, Ross 308 broiler chicks were used. Experiment groups were designed as follows: control; 0.1 % Saccharomyces cerevisiae; 0.2 % Saccharomyces cerevisiae; 0.4 % Saccharomyces cerevisiae. The experimental diets were chemically analyzed according to the methods of the Association of Official Analytical Chemists. Twelve groups were obtained, including three replicates for each experimental group. Each replicated group was comprised of 25 chicks, and thus 75 chicks were placed in each experimental group. After 42 days, broiler chickens were slaughtered. Tibiotarsi were weighed with a digital scale, and the lengths were measured with a digital caliper after the drying process. Cortical areas were measured with the ImageJ Image Processing and Analysis Program. A UTEST Model-7014 tension and compression machine and a Maxtest software were used to determine the bone strength of the tibiotarsus. The severity of the tibial dyschondroplasia lesion was evaluated as 0, +1, +2 and ...
TY - JOUR. T1 - The Saccharomyces cerevisiae gene SDS22 encodes a potential regulator of the mitotic function of yeast type 1 protein phosphatase. AU - MACKELVIE, SARAH H. AU - ANDREWS, PAUL D.. AU - STARK, MICHAEL J. R. PY - 1995/7. Y1 - 1995/7. N2 - In higher eukaryotes, the activity and specificity of the type 1 protein serine-threonine phosphatase (PP1) catalytic subunit is thought to be controlled by its association with a number of regulatory or targeting subunits. Here we describe the characterization of a gene encoding one such potential polypeptide in the yeast Saccharomyces cerevisiae. The gene which we have isolated (termed SDS22) encodes a product with a high degree of sequence identity to the fission yeast sds22 protein, a known regulator of the mitotic function of PP1 in Schizosaccharomyces pombe. Using two different criteria, we have demonstrated that Sds22p and the catalytic subunit of PP1 (Glc7p) interact in yeast cells. We have also generated a temperature-sensitive allele of ...
ARAUJO, Roberta A.C. et al. Monitoring Saccharomyces cerevisiae populations by mtDNA restriction analysis and other molecular typing methods during spontaneous fermentation for production of the artisanal cachaça. Braz. J. Microbiol. [online]. 2007, vol.38, n.2, pp.217-223. ISSN 1517-8382. http://dx.doi.org/10.1590/S1517-83822007000200006.. An ecological study on Saccharomyces cerevisiae populations in spontaneous fermentation has been conducted in three vats of a cachaça distillery in Minas Gerais, Brazil. Ninety-seven yeast isolates were collected at the beginning, the middle and at the end of the production period, and were identified by standard methods. Differentiation between the indigenous S. cerevisiae strains isolated was performed by mitochondrial DNA (mtDNA) restriction analysis, RAPD-PCR, and PCR fingerprint using an intron splice primer. Analysis of the mtDNA restriction profiles revealed 12 different patterns, 11 corresponding to indigenous yeasts (I to XI) and one (XII) to a ...
Yeast co-immunoprecipitation dataset ; Gavin et al, 2006 YBR119W YPL178W YBR119W YML046W YBR119W YKL012W YBR119W YDR235W YBR119W YIL061C YBR119W YDR240C YBR119W YGR013W YBR119W YMR125W YBR152W YER172C YBR152W YMR240C YBR152W YMR288W YBR152W YPL213W YBR152W YIR009W YBR152W YDL043C YBR152W YJL203W YBR152W YDR473C YBR152W YGR091W YBR152W YGR075C YBR152W YPR178W YBR152W YBR055C YBR152W YHR165C YBR152W YDL030W YBR152W YML049C YBR152W YER029C YBR152W YKL173W YBR152W YDL098C YBR152W YOR308C YDL087C YER172C YDL087C YMR288W YDL087C YHR086W YDL087C YER165W YDL087C YML046W YDL087C YKL012W YDL087C YDR235W YDL087C YHR165C YDL087C YML049C YDL087C YER029C YDL087C YLR275W YDL087C YLR147C YDL087C YIL061C YDL087C YKL173W YDL087C YDR240C YDL087C YGR013W YDL087C YMR125W YDL087C YGR162W YDL087C YLR298C YDL175C YJL050W YDL175C YPL190C YDL175C YOL115W YDR235W YHR086W YDR235W YML046W YDR235W YKL012W YDR235W YIL061C YDR235W YGR013W YDR235W YMR125W YDR240C YHR086W YDR240C YML046W YDR240C YDR235W YDR240C YHR165C YDR240C ...
AbstractProtein-metabolite interactions are of crucial importance for all cellular processes but remain understudied. Here, we applied a biochemical approach named PROMIS, to address the complexity of the protein-small molecule interactome in the model yeast Saccharomyces cerevisiae. By doing so, we provide a unique dataset, which can be queried for interactions between 74 small molecules and 3982 proteins using a user-friendly interface available at https://promis.mpimp-golm.mpg.de/yeastpmi/. By interpolating PROMIS with the list of predicted protein-metabolite interactions, we provided experimental validation for 225 binding events. Remarkably, of the 74 small molecules co-eluting with proteins, 36 were proteogenic dipeptides. Targeted analysis of a representative dipeptide, Ser-Leu, revealed numerous protein interactors comprising chaperones, proteasomal subunits, and metabolic enzymes. We could further demonstrate that Ser-Leu binding increases activity of a glycolytic enzyme ...
The Saccharomyces cerevisiae SNF2 gene affects the expression of many diversely regulated genes and has been implicated in transcriptional activation. We report here the cloning and characterization of STH1, a gene that is homologous to SNF2. STH1 is essential for mitotic growth and is functionally distinct from SNF2. A bifunctional STH1-beta-galactosidase protein is located in the nucleus. The predicted 155,914-Da STH1 protein is 72% identical to SNF2 over 661 amino acids and 46% identical over another stretch of 66 amino acids. Both STH1 and SNF2 contain a putative nucleoside triphosphate-binding site and sequences resembling the consensus helicase motifs. The large region of homology shared by STH1 and SNF2 is conserved among other eukaryotic proteins, and STH1 and SNF2 appear to define a novel family of proteins related to helicases. ...
TY - JOUR. T1 - Investigation of steroid receptor function in the budding yeast Saccharomyces cerevisiae. AU - McEwan, I J PY - 1999. Y1 - 1999. N2 - Steroid hormones are small lipophilic molecules that control a wide range of responses in both the developing and adult organism. The actions of these molecules are mediated by soluble receptor proteins that function as hormone-activated transcription factors. The first steroid receptors were expressed in the yeast Saccharomyces cerevisae over 10 years ago, and to date virtually all the classical steroid receptors, together with a number of non-steroid members of the nuclear receptor superfamily, have been expressed in yeast. The ability to reconstitute steroid receptor signalling in yeast cells by co-expression of the receptor protein and a reporter gene driven by the appropriate hormone response element has presented researchers with a powerful model system for investigating receptor action. Tn this review, the use of yeast-based steroid receptor ...
The yeast Saccharomyces cerevisiae is an important eukaryotic workhorse in traditional and modern biotechnology. At present, only a few S. cerevisiae strains have been extensively used as engineering hosts. Recently, an astonishing genotypic and phenotypic diversity of S. cerevisiae was disclosed in natural populations. We suppose that some natural strains can be recruited as superior host candidates in bioengineering. This study engineered a natural S. cerevisiae strain with advantages in inulin utilization to produce ethanol from inulin resources by consolidated bioprocess. Rational engineering strategies were employed, including secretive co-expression of heterologous exo- and endo-inulinases, repression of a protease, and switch between haploid and diploid strains. Results from co-expressing endo- and exo-inulinase genes showed that the extracellular inulinase activity increased 20 to 30-fold in engineered S. cerevisiae strains. Repression of the protease PEP4 influenced cell physiology in late
Saccharomyces cerevisiae is a species of budding yeast. It is perhaps the most useful yeast owing to its use since ancient times in baking and brewing. It is believed that it was originally isolated from the skins of grapes (one can see the yeast as a component of the thin white film on the skins of some dark-colored fruits such as plums; it exists among the waxes of the cuticle). It is one of the most intensively studied eukaryotic model organisms in molecular and cell biology, much like Escherichia coli as the model prokaryote. It is the microorganism behind the most common type of fermentation. Saccharomyces cerevisiae cells are round to ovoid, 5-10 micrometres in diameter. It reproduces by a division process known as budding. It is useful in studying the cell cycle because it is easy to culture, but, as a eukaryote, it shares the complex internal cell structure of plants and animals. S. cerevisiae was the first eukaryotic genome that was completely sequenced. The yeast genome database [1] is ...
The Sec18 protein (Sec18p) of the yeast Saccharomyces cerevisiae has been identified as a component involved in the vesicular transport of proteins through the secretory and endocytotic pathways. Sec18p is a homologue of the mammalian protein NSF which has been shown, using a number of in vitro transport assay systems and affinity purification procedures, to interact with other proteins in a multisubunit protein complex. This work represents two approaches taken with the aim of identifying proteins that interact with Sec18p in the yeast Saccharomyces cerevisiae. Isolation of protein complexes was first attempted by affinity purification of a tagged version of Sec18p. The protein was C-terminally tagged with a protein A moiety from Staphylococcus aureus containing IgG binding domains. It was hoped that the affinity of protein A for IgG Sepharose could be used to isolate protein complexes that formed in vivo with the Sec18p. Although the fusion construct was shown to be active in vivo, specific ...
Under amino acid starvation conditions, the bakers yeast Saccharomyces cerevisiae activates a system called General control of amino acid biosynthesis. Gcn4p, the transcription factor of this system induces the expression of more than 50 genes involved in the different amino acid biosynthetic pathways. In this thesis it could be shown that during simultaneous limitation of amino acids and nitrogen the general control is not activated. More exactly, even a decrease of the Gcn4p activity was detected, which was traced back onto a reduction of the Gcn4 protein amount in the cell. This decrease of the intracellular concentration was caused by translational control of the GCN4 mRNA, which was able to repress even a 2-fold increase of the GCN4 transcription rate. Furthermore during nitrogen starvation conditions no correlation between the stature of eIF-2 phosphorylation and GCN4 expression was observed. For this reason an involvement of the already known mechanism of translation! al regulation of ...
Pulsed electric field (PEF) treatment can be used for non-thermal inactivation of microorganisms. The aim of this paper is to investigate PEF treatment of yeast, Saccharomyces cerevisiae, using three different field waveforms: square; non-oscillating exponential and oscillating exponential. The PEF system used in this paper consists of a pulsed power supply and a parallel-plane metallic electrodes treatment cell located in an air-pressurised chamber. PEF treatment of the yeast was conducted using electric field impulses with magnitudes of 67 kV/cm and 80 kV/cm. The efficacy of the PEF treatment for inactivation of the yeast cells was assessed by comparison of the PEF-treated and untreated yeast populations. Results showed that 3-log10 reduction in the yeast population can be achieved with 100 impulses using all tested waveforms. Amongst all three tested waveforms non-oscillating exponential impulses demonstrated improved PEF performance. The effect of duration of treatment and peak magnitude ...
HOMOLOGOUS recombination is required for the faithful repair of DNA double-strand breaks (DSBs) that arise during normal cellular processes or from exposure of cells to DNA-damaging agents. Central to the process of homologous recombination is the Rad51 protein, which facilitates synapsis and strand invasion into homologous duplex DNA (San Filippo et al. 2008). Rad51 belongs to the RecA family of homologous pairing proteins (Aboussekhra et al. 1992; Basile et al. 1992; Shinohara et al. 1992). Yeast and humans have two RecA homologs: Rad51 and the meiosis-specific Dmc1 (Bishop et al. 1992; San Filippo et al. 2008). In addition, the Saccharomyces cerevisiae RAD55 and RAD57 genes encode proteins with sequence similarity to RecA and Rad51 and are considered to be Rad51 paralogs (Kans and Mortimer 1991; Lovett 1994). Mutation of RAD51, RAD55, or RAD57 confers sensitivity of ionizing radiation (IR) and defects in mitotic and meiotic recombination, indicating that their functions are not redundant ...
TRA1 is an essential gene in Saccharomyces cerevisiae that encodes a 437 kDa protein product. It is a member of a family of key signaling and regulatory molecules that contain a C-terminal phosphatidylinositol-3-kinase (PI3K) domain [1] and is a component of two multisubunit transcriptional regulatory complexes, the SAGA/SLIK and NuA4 complexes, which also contain the histone acetyltransferase enzymes, Gcn5 and Esa1, respectively [2-4]. Tra1 interacts directly with transcriptional activator proteins and is thought to be critical in recruitment of SAGA/SLIK and NuA4 to their target promoters [5-8].. Previously we identified mutations in the C-terminal PI3K domain of Tra1 that showed defects in transcriptional activation, sensitivity to ethanol and the cell wall destabilizing agent calcofluor white and resulted in shortened telomeres [9]. The pattern of changes neither fully mimicked those seen upon disruption of other SAGA/SLIK nor NuA4 components. For example, unlike strains with deletions of ...
During the production of wine and beer, the yeast Saccharomyces cerevisiae can encounter an environment that is deficient in zinc, resulting in a sluggish or a stuck ferment. It has been shown that the Zap1p-transcription factor induces the expression of a regulon in response to zinc deficiency; however, it was evident that a separate regulon was also activated during zinc deficiency in a Zap1p-independent manner. This study discovered the Msn2p and Msn4p (Msn2/4p) transcriptional activator proteins to be an additional control mechanism inducing the stress response during zinc deficiency. Promoter sequence analysis identified the stress response element (STRE) motif, recognized by Msn2/4p, and was significantly enriched in the promoters of genes induced by zinc deficiency. An investigation using genome-wide analyses revealed a distinct regulon consisting of STREcontaining genes whose zinc-responsive expression was abolished in an msn2 msn4 double mutant. An STRE-driven lacZ reporter ...
Yeast cells. Coloured Scanning Electron Micrograph (SEM) of yeast cells, Saccharomyces cerevisiae. This fungus, also known as Bakers or Brewers yeast, consists of single vegetative cells. Some cells can be seen dividing by budding off new cells. Saccharomyces cerevisiae ferments sugar, producing alcohol and carbon dioxide in the process. It has long been used in brewing beer, the production of wine and in baking leavened bread (carbon dioxide causes the dough to rise). Medically, dried Bakers yeast is used as a rich source of vitamin B1, riboflavin and nicotinic acid. Magnification: x125 at 6x7cm size. x200 at 4x5 - Stock Image B250/0646
The effect of yeast (Saccharomyces cerevisiae) on fattening performances of growing cattle is an article from MOJ Ecology & Environmental Sciences for MedCrave Group. The aim of this experiment was to evaluate the yeast on fattening performances of the growing cattle. The experiment was carried out with 179 imported 12-14 months old growing mixed breed bulls (Hereford, Angus, Brangus, and some other crossbreds) that were allocated to control and yeast group according to the breeds and body weight. Experimental diet was formulated with 19 % roughages (alfalfa and wheat straw) containing 13% crude protein. Yeast group was supplemented 40g d-1 live yeast containing 1.23×1011 CFU/g. The study lasted 62 days from May to July. Initial body weight were 393.91±4,43 for control and 395.56±4.45kg for yeast group. After test period, daily gain was similar (1465.85±26.76 vs. 1451.42±34.05g d-1, P|0.05) for the bull receiving the diet without yeast compared to the bulls receiving yeast. Similar results were
Saccharomyces Cerevisiae Yeast Cells Sem Scanning as a 8x6 Glass Mount from CMSP Photo Prints. Fast and safe delivery. Saccharomyces Cerevisiae Yeast Cells. these Microorganisms Fungi are Used to Raise Bread Dough the Yeasts Produce
The Saccharomyces cerevisiae transcription factor Spt20/Ada5 was originally identified by mutations that suppress Ty insertion alleles and by mutations that suppress the toxicity caused by Gal4-VP16 overexpression. Here we present evidence for physical associations between Spt20/Ada5 and three other Spt proteins, suggesting that they exist in a complex. A related study demonstrates that this complex also contains the histone acetyltransferase, Gcn5, and Ada2. This complex has been named SAGA (Spt/Ada/Gcn5 acetyltransferase). To identify functions that genetically interact with SAGA, we have screened for mutations that cause lethality in an spt20Δ/ada5Δ mutant. Our screen identified mutations in SNF2, SIN4, and GAL11. These mutations affect two known transcription complexes: Snf/Swi, which functions in nucleosome remodeling, and Srb/mediator, which is required for regulated transcription by RNA polymerase II. Systematic analysis has demonstrated that spt20Δ/ada5Δ and spt7Δ mutations cause ...
Anhydrobiosis is the state of life when cells get into waterless conditions and gradually cease their metabolism. In this study, we determined the sequence of events in Saccharomyces cerevisiae energy metabolism during processes of dehydration and rehydration. The intensities of respiration and acidification of the medium, the amounts of Phenyldicarbaundecaborane (PCB-) bound to yeast membranes, and the capabilities of cells to accumulate K+ were assayed using electrochemical monitoring system, and intracellular content of ATP was measured using bioluminescence assay. Mesophilic, semi-resistant to desiccation S. cerevisiae strain 14 and thermotolerant, very resistant to desiccation S. cerevisiae strain 77 cells were compared. After 22 h of drying it was possible to restore the respiration activity of very resistant to desiccation strain 77 cells, especially when glucose was available. PCB- binding also indicated considerably higher metabolic activity of dehydrated S. cerevisiae strain 77 cells.
Saccharomyces cerevisiae sudah sejak lama digunakan sebagai starter fermentasi pembuatan roti dan minuman beralkohol. Dalam buku ini, Saccharomyces crervisiae dimanfaatkan sebagai agensia modifikasi dalam pengolahan pangan, kemampuan S. cerevisiae dalam merombak komponen pangan, produk metabolit yang dihasilkan oleh S. cerevisiae, modifikasi terhadap perubahan sifat beberapa produk pangan oleh S. cerevisiae seperti tapioka, tempe, dan modifikasi fermentasi kakao. Pengertian dasar mengenai khamir perlu dipahami oleh mahasiswa yang khususnya mempelajari mikrobiologi pangan, mikrobiologi industri dan teknologi pangan. S.cerevisiae adalah khamir ...
Budding Yeast: Saccharomyces cerevisiae Saccharomyces cerevisiae, the budding yeast, is the common yeast used in baking (bakers yeast) and brewing (brewers
The production of bio-based chemicals, fuels, pharmaceuticals and food additives by microbial fermentation is a rapidly growing field. There is an increasing demand for efficient cell factories that enable the production of biofuels and biochemicals from renewable resources at low and competitive cost. The knowledge of genetics, physiology, biochemistry and large-scale fermentation of bakers yeast Saccharomyces cerevisiae, combined with the advent of genome engineering and recombinant DNA technology makes it a preferred host for many industrial bio-based applications, ranging from biofuels and bulk chemicals to nutraceuticals and pharmaceuticals [1-8]. Furthermore, S. cerevisiae has the advantage of being easy to manipulate genetically with a range of established cloning and vector systems [6, 9].. Production organisms with multi-enzyme pathways often require precise control of the expression level of the associated genes [2, 5, 10]. Besides regulating promoter strength, the copy number of ...
Bakers yeast Saccharomyces cerevisiae is one of the most important and widely used cell factories for recombinant protein production. Many strategies have been applied to engineer this yeast for improving its protein production capacity, but productivity is still relatively low, and with increasing market demand, it is important to identify new gene targets, especially targets that have synergistic effects with previously identified targets. Despite improved protein production, previous studies rarely focused on processes associated with intracellular protein retention. Here we identified genetic modifications involved in the secretory and trafficking pathways, the histone deacetylase complex, and carbohydrate metabolic processes as targets for improving protein secretion in yeast. Especially modifications on the endosome-to-Golgi trafficking was found to effectively reduce protein retention besides increasing protein secretion. Through combinatorial genetic manipulations of several of the newly
Saccharomyces cerevisiae ATCC ® 201390D-5™ Designation: Genomic DNA from Saccharomyces cerevisiae Strain BY4743 (ATCC ® 201390™) Application:
Saccharomyces cerevisiae ATCC ® 201389D-5™ Designation: Genomic DNA from Saccharomyces cerevisiae Strain BY4742 (ATCC ® 201389™) Application:
TY - JOUR. T1 - Influence of cell surface characteristics on adhesion of Saccharomyces cerevisiae to the biomaterial hydroxylapatite. AU - White, Jane S.. AU - Walker, Graeme M.. PY - 2011/2. Y1 - 2011/2. N2 - The influence of the physicochemical properties of biomaterials on microbial cell adhesion is well known, with the extent of adhesion depending on hydrophobicity, surface charge, specific functional groups and acid-base properties. Regarding yeasts, the effect of cell surfaces is often overlooked, despite the fact that generalisations may not be made between closely related strains. The current investigation compared adhesion of three industrially relevant strains of Saccharomyces cerevisiae (M-type, NCYC 1681 and ALY, strains used in production of Scotch whisky, ale and lager, respectively) to the biomaterial hydroxylapatite (HAP). Adhesion of the whisky yeast was greatest, followed by the ale strain, while adhesion of the lager strain was approximately 10-times less. According to ...
The Pumilio family (PUF) proteins are conserved among the eukaryotes (42). They bind to specific sequences in the 3′ untranslated region (3′UTR) of target transcripts via their conserved and characteristic PUF domain and thereby inhibit the stability or translatability of these target mRNAs (32, 50). Indeed, the PUF domain appears sufficient for PUF proteins to affect their target transcripts (32, 50). Five PUF proteins, Puf1p to Puf5p, were thought to exist in the budding yeast Saccharomyces cerevisiae (37, 49). A sixth, Puf6p, has recently been reported (9). None are essential (9, 37, 49). One of the yeast PUF proteins, Mpt5p, also known as Htr1p (23), Puf5p (37), or Uth4p (20), promotes replicative life span (3, 20, 21), the number of generations a virgin daughter cell can undergo before becoming senescent. Mpt5p is a robust regulator of ageing, since it also affects life span in a long-lived genetic background (17).. In addition to displaying a short replicative life span, mutants ...
TY - JOUR. T1 - Dynamic Effects Related to Steady-State Multiplicity in Continous Saccharomyces Cerevisiae Cultivations. AU - Lei, Frede. AU - Olsson, Lisbeth. AU - Jørgensen, Sten Bay. PY - 2004. Y1 - 2004. N2 - The behavioral differences between chemostat and productostat cultivation of aerobic glucose-limited Saccharomyces cerevisiae were investigated. Three types of experiments were conducted: a chemostat, where the dilution rate was shifted up or down in stepwise manner; and a productostat, with either stepwise changed or a rampwise increased ethanol setpoint, i.e., an accelero-productostat. The transient responses from chemostat and productostat experiments were interpreted using a simple metabolic flux model. In a productostat it was possible to obtain oxido-reductive steady states at dilution rates far below D-crit due to a strong repression of the respiratory system. However, these steady states could not be obtained in a chemostat, since a dilution rate shift-down from an ...
TY - CHAP. T1 - Lipids and membranes in Saccharomyces cerevisiae.. AU - Schweizer, Michael. PY - 1999. Y1 - 1999. M3 - Chapter. SP - 79. EP - 155. BT - In The Metabolism & Molecular Physiology of Saccharomyces cerevisiae. Eds. J R Dickinson & M Schweizer. Taylor & Francis, London. ER - ...
To grow, eukaryotic cells must expand by inserting glycerolipids, sphingolipids, sterols, and proteins into their plasma membrane, and maintain the proper levels and bilayer distribution. A fungal cell must coordinate growth with enlargement of its cell wall. In Saccharomyces cerevisiae, a plasma membrane‐localized protein kinase complex, Target of Rapamicin (TOR) complex‐2 (TORC2) (mammalian ortholog is mTORC2), serves as a sensor and masterregulator of these plasma membrane‐ and cell wall‐associated events by directly phosphorylating and thereby stimulating the activity of two types of effector protein kinases: Ypk1 (mammalian ortholog is SGK1), along with a paralog (Ypk2); and, Pkc1 (mammalian ortholog is PKN2/PRK2). Ypk1 is a central regulator of pathways and processes required for plasma membrane lipid and protein homeostasis, and requires phosphorylation on its T‐loop by eisosome‐associated protein kinase Pkh1 (mammalian ortholog is PDK1) and a paralog (Pkh2). For cell survival under
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The structure of a polysaccharide consisting of D-glucose isolated from the cell-wall of active dry bakers yeast (Saccharomyces cerevisiae) was investigated by using methylation analysis, periodate oxidation, mass spectrometry, NMR spectroscopy, and enzymic hydrolysis, as a new approach in determination of structures. The main structural feature of the polysaccharide deduced on the basis of the obtained results is a linear chain of (1→3)-linked β-D-glucopyranoses, a part of which is substituted through the positions O-6. The side units or groups are either a single D-glucopyranose or (1→3)-β-oligoglucosides, linked to the main chaing through (1→6)-glucosidic linkages. The low optical rotation as well as the 13C-NMR and FTIR spectra suggest that the glycosidic linkages are in the β-D-configuration ...
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We have isolated STN1, an essential Saccharomyces cerevisiae gene, as a suppressor of the cdc13-1 mutation. A synthetic lethal interaction between a temperature-sensitive mutant allele of STN1, stn1-13, and cdc13-1 was observed. Stn1 and Cdc13 proteins displayed a physical interaction by two-hybrid analysis. As shown previously for cdc13-1, stn1-13 cells at the restrictive temperature accumulate single-stranded DNA in subtelomeric regions of the chromosomes, but to a lesser extent than cdc13-1 cells. In addition, both Cdc13 and Stn1 were found to be involved in the regulation of telomere length, mutations in STN1 or CDC13 conferring an increase in telomere size. Loss of Stn1 function activated the RAD9 and MEC3 G2/M checkpoints, therefore confirming that DNA damage is generated. We propose that Stn1 functions in telomere metabolism during late S phase in cooperation with Cdc13 ...
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Base Sequence, DNA Polymerase II/chemistry, DNA Polymerase III/*chemistry/metabolism, DNA Replication, Molecular Sequence Data, Saccharomyces cerevisiae/enzymology/*genetics, Saccharomyces cerevisiae Proteins/*chemistry ...
All alcohols and spirits such as beer need yeast for the purpose of fermentation and leading breweries understand that getting top quality beer with saccharomyces cerevisiae yeast is the only way to happily placate parched throats of enthusiastic drinkers all around the globe.. All types of alcohols and also spirits like beer, wines, whiskey, rum, vodka, and so on have got diverse alcohol strengths. Different types of yeast as well can merely ferment as well as survive within a variety of alcohols, and are additionally limited by temperature distillersyeast. Thus alcoholic beverage manufacturing involving the production of vodka cannot use yeast suitable for lower alcohol potency yeasts such as wine yeast or even other forms of yeast such as saccharomyces cerevisiae, which is essentially used for brewing beer.. The saccharomyces cerevisiae yeast is actually belonging to the fungi family similar to its other cousins which ferment various other kinds of alcohols. This particular yeast is often ...
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Understanding how new biochemical pathways evolve in a sexually reproducing population is a complex and largely unanswered question. We have successfully evolved a novel biochemical pathway in yeast using a sex based population approach.. For over 30 years, wild type Saccharomyces has been widely reported to not grow on xylose at all, but we discovered that most strains can grow, albeit at almost undetectable rates. A mass mated starting population of Saccharomyces cerevisiae strains was evolved under selection on Xylose Minimal Media (XMM) with forced sexual mating every ~two months for 1463 days. This produced a population that could grow on xylose as a sole carbon source. Initial studies show the xylose growth trait is quantitative and presumably governed by many genes. To investigate the evolution of the xylose phenotype, a xylose utilising strain MBG11a was isolated. MBG11a was sequenced with PacBio RSII long read sequencing at the Ramaciotti Centre for Genomics. A high quality complete ...
Background and objectivePineapple peels contain significant quantities of carbohydrates, which can be used as cheap raw materials for production of commercially important products through fermentation. The aim of this study was to use this feed stock for the cultivation of Saccharomyces cerevisiae NCDC 364 and its use as single cell protein.Material and methodsThe single cell protein was produced using discarded pineapple peels and Saccharomyces cerevisiae NCDC 364. Optimization of bioprocess variables (temperature, pH, incubation period, carbon source and nitrogen source) affecting single cell protein production was carried out using classical one factor at a time approach. The harvested cells from optimized media were screened for amino acid content using high-performance thin-layer chromatography.Results andconclusionThe Saccharomyces cerevisiae NCDC 364 produced maximum single cell protein in pineapple peel based media, compared to non-optimized media. The one factor at a time approach showed
New DNA Sequences ======================= AF200324 AF200324 579bp DNA PLN 03-FEB-2000 Saccharomyces cerevisiae Tim18p (TIM18) gene, complete cds; nuclear gene for mitochondrial product. TIM18; Tim18p. =========== Updated Sequence Features/Annotations ============= D37948 D37948 1777bp DNA PLN 01-FEB-2000 Saccharomyces cerevisiae DNA for F1F0-ATPase alpha subunit precursor, complete cds. ATP1; F1F0-ATPase alpha subunit precursor; F1F0-ATPase alpha subunit. D37949 D37949 1778bp DNA PLN 01-FEB-2000 Saccharomyces cerevisiae DNA for defective F1F0-ATPase alpha subunit precursor, complete cds. F1F0-ATPase alphadefective F1F0-ATPase alpha subunit precursor; defective F1F0-ATPase alpha subunit. YSC9SF D00347 870bp DNA PLN 01-FEB-2000 Saccharomyces cerevisiae 9kDa stabilizing factor. ATP synthase; ATP synthase inhibitor protein; mitochondria; 9kDa stabilizing factor mature protein. YSCCOF D13230 2005bp DNA PLN 01-FEB-2000 Saccharomyces cerevisiae COF1 gene for cofilin, complete cds. cofilin; COF1. ...
TY - JOUR. T1 - A dependent pathway of gene functions leading to chromosome segregation in saccharomyces cerevisiae. AU - Wood, John S.. AU - Hartwell, Leland H.. N1 - Copyright: Copyright 2017 Elsevier B.V., All rights reserved.. PY - 1982/9/1. Y1 - 1982/9/1. N2 - Methyl-benzimidazole-2-ylcarbamate (MBC) inhibits the mitotic cell cycle of Saccharomyces cerevisiae at a stage subsequent to DNA synthesis and before the completion of nuclear division (Quinlan, R. A., C. I. Pogson, and K. Gull, 1980, J. Cell 5ci., 46: 341-352). The step in the cell cycle that is sensitive to M8C inhibition was ordered in reciprocal shift experiments with respect to the steps catalyzed by cdc gene products. Execution of the CDC7 step is required for the initiation of DNA synthesis and for completion of the MBC-sensitive step. Results obtained with mutants (cdc2, 6, 8, 9, and 21) defective in DNA replication and with an inhibitor of DNA replication (hydroxyurea) suggest that some DNA replication is required for ...
It is essential when studying the circadian rhythm in cells to be able to effectively stop them in time. In this experiment, we tested what would be the most successful killing agent on Saccharomyces cerevisiae. Six different agents were tested at different concentrations and amounts. After the S. cerevisiae was added to the test tube containing the agent, it was streaked on a plate after 5 and 10 minutes. The plates were incubated and then checked for growth. Ethanol was the most efficient killing agent. After an effective killing agent is determined, it can be used in further experiments measuring Gapdehydrogenase activity using a colorimetric assay to examine the circadian rhythm in Saccharomyces cerevisiae. Gapdehydrogenase results will also be presented.
Autophagy is an intracellular process responsible for the degradation and recycling of cytoplasmic components. It selectively removes harmful cellular material and enables the cell to survive starvation by mobilizing nutrients via the bulk degradation of cytoplasmic components. While research over the last decades has led to the discovery of the key factors involved in autophagy, the pathway is not yet completely understood. The first studies of autophagy on a molecular level were conducted in the yeast Saccharomyces cerevisiae. Building up on these studies, many homologs have been found in higher eukaryotes. Yeast remains a highly relevant model organism for studying autophagy, with a wide range of established methods to elucidate the molecular details of the autophagy pathway. In this review, we provide an overview of methods to study both selective and bulk autophagy, including intermediate steps in the yeast Saccharomyces cerevisiae. We compare different assays, discuss their advantages and
Abstract: The objective of this study was to select three strains of probiotic Saccharomyces cerevisiae and to evaluate the effect of S. cerevisiae and rumen bacteria isolate (MR4) supplementation and their combination on rumen fermentability and rumen microbial population. Experiment 1 was designed in a 4 x 5 factorial randomized block design with 3 replications. The first factor was S. cerevisiae strain consisted of control treatment (without S. cerevisiae supplementation), NBRC 10217, NRRL Y 567 and NRRL 12618, and the second factor was incubation time consisted of 0, 1, 2, 3, and 4 h. Ration was basal ration for feedlot with forage to concentrate ratio (F:C)= 60:40. Dosage of each treatment with S. cerevisiae was 5 x 1010 cfu/kg ration. Experiment 2 was designed in randomized block design with 4 treatments: P0= basal ration of feedlot; P1= P0 + S. cerevisiae; P2= P0 + MR4 isolate (5 x 107 cfu/kg ration); P3= P0 + S. cerevisiae and MR4 isolate. The result of experiment 1 showed that ...
Article Saccharomyces cerevisiae afr1 protein is a protein phosphatase 1/glc7-targeting subunit that regulates the septin cytoskeleton during mating. Glc7, the type1 serine/threonine phosphatase in the yeast Saccharomyces cerevisiae, is targeted by a...
TY - CHAP. T1 - Delta integration CRISPR-Cas (Di-CRISPR) in saccharomyces cerevisiae. AU - Shi, Shuobo. AU - Liang, Youyun. AU - Ang, Ee Lui. AU - Zhao, Huimin. PY - 2019/1/1. Y1 - 2019/1/1. N2 - Despite the advances made in genetic engineering of Saccharomyces cerevisiae, the multicopy genomic integration of large biochemical pathways remains a challenge. Here, we developed a Di-CRISPR (delta integration CRISPR-Cas) platform based on cleavage of the delta sites by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated systems (Cas) to enable unprecedented high-efficiency, multicopy, markerless integrations of large biochemical pathways into the S. cerevisiae genome. Detailed protocols are provided on the entire workflow which includes pDi-CRISPR plasmid and donor DNA construction, Di-CRISPR-mediated integration and analysis of integration efficiencies and copy numbers through flow cytometry and quantitative polymerase chain reaction (qPCR).. AB - Despite the ...
TY - JOUR. T1 - The effect of phosphate accumulation on metal ion homeostasis in Saccharomyces cerevisiae. AU - Rosenfeld, Leah. AU - Reddi, Amit R.. AU - Leung, Edison. AU - Aranda, Kimberly. AU - Jensen, Laran T.. AU - Culotta, Valeria C.. PY - 2010/9/1. Y1 - 2010/9/1. N2 - Much of what is currently understood about the cell biology of metals involves their interactions with proteins. By comparison, little is known about interactions of metals with intracellular inorganic compounds such as phosphate. Here we examined the role of phosphate in metal metabolism in vivo by genetically perturbing the phosphate content of Saccharomyces cerevisiae cells. Yeast pho80 mutants cannot sense phosphate and have lost control of phosphate uptake, storage, and metabolism. We report here that pho80 mutants specifically elevate cytosolic and nonvacuolar levels of phosphate and this in turn causes a wide range of metal homeostasis defects. Intracellular levels of the hard-metal cations sodium and calcium ...
A new aspartic protease from Saccharomyces cerevisiae, with a high degree of similarity with yapsin 1 and yapsin 2 and a specificity for basic residue cleavage sites of prohormones, has been cloned. This enzyme was named yapsin 3. Expression of a C-terminally truncated non-membrane anchored yapsin 3 in yeast yielded a heterogeneous protein between 135-200 kDa which, upon treatment with endoglycosidase H, migrated as a 60 kDa form. Amino-acid analysis of the N-terminus of expressed yapsin 3 revealed two different N-terminal residues, serine-48 and phenylalanine-54, which followed a dibasic and a monobasic residue respectively. Cleavage of several prohormones by non-anchored yapsin 3 revealed a specificity distinct from that of yapsin 1.. ...
Saccharomyces Cerevisiae Yeast Cells Sem Scanning as a A2 (42x59 cm) Fine Art Print from CMSP Photo Prints. Fast and safe delivery. Saccharomyces Cerevisiae Yeast Cells. these Microorganisms Fungi are Used to Raise Bread Dough the Yeasts Produce
MIG1 overexpression causes flocculation in Saccharomyces cerevisiae. An important role of glutathione and gamma-glutamyltranspeptidase in the supply of growth requirements during nitrogen starvation of the yeast Saccharomyces cerevisiae
TY - JOUR. T1 - Magnesium as a stress-protectant for industrial strains of saccharomyces cerevisiae. AU - Walker, Graeme M.. PY - 1998. Y1 - 1998. N2 - During brewery fermentations, individual yeast cells may be confronted with a variety of environmental stresses that impair yeast growth and fermentative metabolism. An understanding of the stress physiology of industrial yeasts is therefore important in order to counteract deleterious effects of stress on fermentation and, ultimately, product quality. The present study describes the influence of magnesium ions in preventing cell death caused by temperature shock and ethanol toxicity in Saccharomyces cerevisiae yeast strains employed in brewing, distilling, and wine fermentations. Results obtained show that, by increasing the extracellular availability of magnesium ions, physiological protection may be conferred on temperature- and ethanol-stressed yeast cells with respect to culture viability and growth. This practical approach is envisaged to ...
The Saccharomyces cerevisiae SIS1 gene was identified as a high copy number suppressor of the slow growth phenotype of strains containing mutations in the SIT4 gene, which encodes a predicted serine/threonine protein phosphatase. The SIS1 protein is similar to bacterial dnaJ proteins in the amino-terminal third and carboxyl-terminal third of the proteins. In contrast, the middle third of SIS1 is not similar to dnaJ proteins. This region of SIS1 contains a glycine/methionine-rich region which, along with more amino-terminal sequences, is required for SIS1 to associate with a protein of apparent molecular mass of 40 kD. The SIS1 gene is essential. Strains limited for the SIS1 protein accumulate cells that appear blocked for migration of the nucleus from the mother cell into the daughter cell. In addition, many of the cells become very large and contain a large vacuole. The SIS1 protein is localized throughout the cell but is more concentrated at the nucleus. About one-fourth of the SIS1 protein is ...
en] Time-series of high throughput gene sequencing data intended for gene regulatory network (GRN) inference are often short due to the high costs of sampling cell systems. Moreover, experimentalists lack a set of quantitative guidelines that prescribe the minimal number of samples required to infer a reliable GRN model. We study the temporal resolution of data vs.quality of GRN inference in order to ultimately overcome this deficit. The evolution of a Markovian jump process model for the Ras/cAMP/PKA pathway of proteins and metabolites in the G1 phase of the Saccharomyces cerevisiae cell cycle is sampled at a number of different rates. For each time-series we infer a linear regression model of the GRN using the LASSO method. The inferred network topology is evaluated in terms of the area under the precision-recall curve (AUPR). By plotting the AUPR against the number of samples, we show that the trade-off has a, roughly speaking, sigmoid shape. An optimal number of samples corresponds to values ...
en] Time-series of high throughput gene sequencing data intended for gene regulatory network (GRN) inference are often short due to the high costs of sampling cell systems. Moreover, experimentalists lack a set of quantitative guidelines that prescribe the minimal number of samples required to infer a reliable GRN model. We study the temporal resolution of data vs.quality of GRN inference in order to ultimately overcome this deficit. The evolution of a Markovian jump process model for the Ras/cAMP/PKA pathway of proteins and metabolites in the G1 phase of the Saccharomyces cerevisiae cell cycle is sampled at a number of different rates. For each time-series we infer a linear regression model of the GRN using the LASSO method. The inferred network topology is evaluated in terms of the area under the precision-recall curve (AUPR). By plotting the AUPR against the number of samples, we show that the trade-off has a, roughly speaking, sigmoid shape. An optimal number of samples corresponds to values ...
The yeast assimilatory sulfate reductase is a complex enzyme that is responsible for conversion of sulfite into sulfide. To obtain information on the nature of this enzyme, we isolated and sequenced the MET10 gene of Saccharomyces cerevisiae and a divergent MET10 allele from Saccharomyces carlsbergensis. The polypeptides deduced from the identically sized open reading frames (1,035 amino acids) of both MET10 genes have molecular masses of around 115 kDa and are 88% identical to each other. The transcript of S. cerevisiae MET10 has a size comparable to that of the open reading frame and is transcriptionally repressed by methionine in a way similar to that seen for other MET genes of S. cerevisiae. Distinct homology was found between the putative MET10-encoded polypeptide and flavin-interacting parts of the sulfite reductase flavoprotein subunit (encoded by cysJ) from Escherichia coli and several other flavoproteins. A significant N-terminal homology to pyruvate flavodoxin oxidoreductase (encoded ...
TY - JOUR. T1 - Methylation of translation-associated proteins in Saccharomyces cerevisiae. T2 - Identification of methylated lysines and their methyltransferases. AU - Couttas, Timothy A.. AU - Raftery, Mark J.. AU - Padula, Matthew P.. AU - Herbert, Ben R.. AU - Wilkins, Marc R.. PY - 2012/4. Y1 - 2012/4. N2 - This study aimed to identify sites of lysine methylation in Saccharomyces cerevisiae and the associated methyltransferases. Hexapeptide ligand affinity chromatography was used to normalize the abundance levels of proteins in whole cell lysate. MS/MS, in association with antibody-based detection, was then used to identify lysine methylated proteins and the precise sites of modification. Lysine methylation was found on the proteins elongation factor (EF) 1-α, 2, and 3A, as well as ribosomal proteins 40S S18-A/B, 60S L11-A/B, L18-A/B, and L42-A/B. Precise sites were mapped in all cases. Single-gene knockouts of known and putative methyltransferase(s), in association with MS/MS, showed that ...
Saccharomyces cerevisiae mutants deficient in superoxide dismutase genes (sod1∆, sod2∆and the double mutant) were subjected to H2O2 stress in the stationary phase. The highest sensitivity was observed in the sod2∆mutant, while the sod1∆sod2∆double mutant was not sensitive. sod mutants had lower catalase activity (44%) than wildtype cells, independent of H2O2 stress. Untreated cells of sod1∆sod2∆ double mutants showed increased glutathione peroxidase activity (126%), while sod1∆had lower activity (77%) than the wild type. Glutathione levels in sod1∆were increased (200-260%) after exposure to various H2O2 concentrations. In addition, the highest malondialdehyde levels could be observed without H2O2 treatment in sod1∆ (167%) and sod2∆(225%) mutants. In contrast, the level of malondialdehyde in the sod1∆sod2∆double mutant was indistinguishable from that of the wild type. These results suggest that resistance to H2O2 by sod1∆sod2∆cells depends on the ...
The recently sequenced genome of the filamentous fungus Ashbya gossypii revealed remarkable similarities to that of the budding yeast Saccharomyces cerevisiae both at the level of homology and synteny (conservation of gene order). Thus, it became possible to reinvestigate the S. cerevisiae genome in the syntenic regions leading to an improved annotation. We have identified 23 novel S. cerevisiae open reading frames (ORFs) as syntenic homologs of A. gossypii genes; for all but one, homologs are present in other eukaryotes including humans. Other comparisons identified 13 overlooked introns and suggested 69 potential sequence corrections resulting in ORF extensions or ORF fusions with improved homology to the syntenic A. gossypii homologs. Of the proposed corrections, 25 were tested and confirmed by resequencing. In addition, homologs of nearly 1,000 S. cerevisiae ORFs, presently annotated as hypothetical, were found in A. gossypii at syntenic positions and can therefore be considered as authentic genes.
The yeast Saccharomyces cerevisiae strain W303 synthesizes in the early logarithmic phase of growth dolichols of 14-18 isoprene residues. The analysis of the polyisoprenoids present in the stationary phase revealed an additional family which proved to be also dolichols but of 19-24 isoprene residues, constituting 39% of the total dolichols. The transfer of early logarithmic phase cells to a starvation medium lacking glucose or nitrogen resulted in the synthesis of the longer chain dolichols. The additional family of dolichols represented 13.8% and 10.3% of total dolichols in the glucose and nitrogen deficient media, respectively. The level of dolichols in yeast cells increased with the age of the cultures. Since both families of dolichols are present in stationary phase cells we postulate that the longer chain dolichols may be responsible for the physico-chemical changes in cellular membranes allowing yeast cells to adapt to nutrient deficient conditions to maintain long-term viability ...
We present an example of expression and purification of a biologically active G-protein coupled receptor (GPCR) from yeast. An expression vector was constructed to encode the Saccharomyces cerevisiae GPCR a-factor receptor (Ste2p, the STE2 gene product) containing a 9-amino acid sequence of rhodopsin that served as an epitope/affinity tag. In the construct, two glycosylation sites and two cysteine residues were removed to aid future structural and functional studies. The receptor was expressed in yeast cells and was detected as a single band in a western blot indicating the absence of glycosylation. Ligand binding and signaling assays of the epitope-tagged mutated receptor showed it maintained the full wild-type biological activity. For extraction of Ste2p, yeast membranes were solubilize with 0.5% n-dodecyl maltoside (DM). Approximately 120 mu g of purified a-factor receptor was obtained per liter of culture by single-step affinity chromatography using a monoclonal antibody to the rhodopsin ...
Water-insoluble glucan was isolated from the bakers yeast Saccharomyces cerevisiae. The yeast cells were treated with alkali and the residue then with acid. Chemical and NMR (1D and 2D) analyses showed that a linear (1→3)-β-glucan was purified that was not contaminated with other carbohydrates, proteins or phenolic compounds. The effects of the glucan on wound healing were assessed in human venous ulcers by histopathological analysis after 30 days of topical treatment. (1→3)-β-glucan enhanced ulcer healing and increased epithelial hyperplasia, as well as increased inflammatory cells, angiogenesis and fibroblast proliferation. In one patient who had an ulcer that would not heal for over 15 years, glucan treatment caused a 67.8% decrease in the area of the ulcer. This is the first study to investigate the effects of (1→3)-β-glucan on venous ulcer healing in humans; our findings suggest that this glucan is a potential natural biological response modifier in wound ...
The communication reports the cloning, sequencing, and analysis of the RPS3 gene from yeast, which codes for the ribosomal protein YS3. Sequence analyses of a 2.45 kb DNA fragment revealed an open reading frame with the potential to code for a 240 amino-acid long protein. The first 20 amino acids display a 90% identity to a 20 amino-acid long protein sequence of yeast ribosomal protein S3, that was obtained by protein sequencing of purified yeast ribosomal proteins. The promoter region of the RPS3 gene contains several upstream conserved sequence elements (UASrpg, T-rich region) that usually regulate transcription of ribosomal protein genes. Northern blot experiments demonstrate that this ORF is transcribed into an approximately 900 nt long mRNA. The major start site of transcription is located near position -20. The RPS3 gene is a single copy gene in yeast. Its disruption yields non viable haploid spores of Saccharomyces cerevisiae.
During pretreatment of lignocellulose raw material, compounds that severely inhibit microbial activity including Saccharomyces cerevisiae strains are released [1]. These compounds, which include furaldehydes and weak organic acids, inhibit yeast metabolism and affect yeast viability and, as a consequence, reduces the overall productivity of an ethanol production process [2]. Elucidation of the molecular mechanisms behind inhibition can suggest new strategies to prevent the inhibitory effect. In the present study, the possible effect on the plasma membrane in S. cerevisiae is studied as a response to inhibitors present in lignocellulose raw material. A comparative lipidomic profiling will be carried out on S. cerevisiae cultured in the absence and presence of lignocellulose inhibitors. LC-CAD and GC-MS will be used to extensively characterize the composition of the plasma membrane. Changes in membrane composition will be correlated with the presence of specific inhibitors. References 1. Palmqvist E, Hahn
Every cell division in budding yeast is inherently asymmetric and counts on the correct positioning of the mitotic spindle along the mother-daughter polarity axis for faithful chromosome segregation. A surveillance mechanism named the spindle position checkpoint (SPOC), monitors the orientation of the mitotic spindle and prevents cells from exiting mitosis when the spindle fails to align along the mother-daughter axis. SPOC is essential for maintenance of ploidy in budding yeast and similar mechanisms might exist in higher eukaryotes to ensure faithful asymmetric cell division. Here, we review the current model of SPOC activation and highlight the importance of protein localization and phosphorylation for SPOC function.
DNA replication forks that are stalled by DNA damage activate an S-phase checkpoint that prevents irreversible fork arrest and cell death. The increased cell death caused by DNA damage in budding yeast cells lacking the Rad53 checkpoint protein kinase is partially suppressed by deletion of the gene. Using a whole-genome sequencing approach, we identified two additional genes, and , whose mutation can also partially suppress this DNA damage sensitivity. We provide evidence that and act in a common pathway, which is distinct from the pathway. Analysis of additional mutants indicates that suppression works through the loss of the Rpd3L histone deacetylase complex. Our results suggest that the loss or absence of histone acetylation, perhaps at stalled forks, may contribute to cell death in the absence of a functional checkpoint. ...
In Saccharomyces cerevisiae the activity for the lactate-proton symporter is dependent on JEN1 gene expression. Pichia pastoris was transformed with an integrative plasmid containing the JEN1 gene. After 24 h of methanol induction, Northern and Western blotting analyses indicated the expression of JEN1 in the transformants. Lactate permease activity was obtained in P. pastoris cells with a Vmax of 2.1 nmol·s−1·mg of dry weight−1. Reconstitution of the lactate permease activity was achieved by fusing plasma membranes of P. pastoris methanol-induced cells with Escherichia coli liposomes containing cytochrome c oxidase, as proton-motive force. These assays in reconstituted heterologous P. pastoris membrane vesicles demonstrate that S. cerevisiae Jen1p is a functional lactate transporter. Moreover, a S. cerevisiae strain deleted in the JEN1 gene was transformed with a centromeric plasmid containing JEN1 under the control of the glyceraldehyde-3-phosphate dehydrogenase constitutive promotor. ...
Effect of the msb3msb4 double mutation on the intracellular pool of purine nucleotides in the yeast Saccharomyces cerevisiae: application to the study of the biological activity of the oncogenic human protein oncTre210p ...
TY - JOUR. T1 - Functional profiling of the Saccharomyces cerevisiae genome. AU - Giaever, Guri. AU - Chu, Angela M.. AU - Ni, Li. AU - Connelly, Carla. AU - Riles, Linda. AU - Véronneau, Steeve. AU - Dow, Sally. AU - Lucau-Danila, Ankuta. AU - Anderson, Keith. AU - André, Bruno. AU - Arkin, Adam P.. AU - Astromoff, Anna. AU - El Bakkoury, Mohamed. AU - Bangham, Rhonda. AU - Benito, Rocio. AU - Brachat, Sophie. AU - Campanaro, Stefano. AU - Curtiss, Matt. AU - Davis, Karen. AU - Deutschbauer, Adam. AU - Entian, Karl Dieter. AU - Flaherty, Patrick. AU - Foury, Francoise. AU - Garfinkel, David J.. AU - Gerstein, Mark. AU - Gotte, Deanna. AU - Güldener, Ulrich. AU - Hegemann, Johannes H.. AU - Hempel, Svenja. AU - Herman, Zelek. AU - Jaramillo, Daniel F.. AU - Kelly, Diane E.. AU - Kelly, Steven L.. AU - Kötter, Peter. AU - LaBonte, Darlene. AU - Lamb, David C.. AU - Lan, Ning. AU - Liang, Hong. AU - Liao, Hong. AU - Liu, Lucy. AU - Luo, Chuanyun. AU - Lussier, Marc. AU - Mao, Rong. AU - ...
One of the most crucial tasks for a cell to ensure its long term survival is preserving the integrity of its genetic heritage via maintenance of DNA structure and sequence. While the DNA damage response in the yeast Saccharomyces cerevisiae, a model eukaryotic organism, has been extensively studied, much remains to be elucidated about how the organism senses and responds to different types and doses of DNA damage. We have measured the global transcriptional response of S. cerevisiae to multiple doses of two representative DNA damaging agents, methyl methanesulfonate (MMS) and gamma radiation. Hierarchical clustering of genes with a statistically significant change in transcription illustrated the differences in the cellular responses to MMS and gamma radiation. Overall, MMS produced a larger transcriptional response than gamma radiation, and many of the genes modulated in response to MMS are involved in protein and translational regulation. Several clusters of coregulated genes whose responses varied
Lignocellulosic biomass yields after hydrolysis, besides the hexose D-glucose, D-xylose, and L-arabinose as main pentose sugars. In second generation bioethanol production utilizing the yeast Saccharomyces cerevisiae, it is critical that all three sugars are co-consumed to obtain an economically feasible and robust process. Since S. cerevisiae is unable to metabolize pentose sugars, metabolic pathway engineering has been employed to introduce the respective pathways for D-xylose and L-arabinose metabolism. However, S. cerevisiae lacks specific pentose transporters, and these sugars enter the cell with low affinity via glucose transporters of the Hxt family. Therefore, in the presence of D-glucose, utilization of D-xylose and L-arabinose is poor as the Hxt transporters prefer D-glucose. To solve this problem, heterologous expression of pentose transporters has been attempted but often with limited success due to poor expression and stability, and/or low turnover. A more successful approach is the ...
Read Expression of the Drosophila melanogaster limk1 gene 3′-UTRs mRNA in yeast Saccharomyces cerevisiae, Russian Journal of Genetics on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.