The reaction of cytochrome oxidase with oxygen in the fission yeast Schizosaccharomyces pombe 972h-. Studies at subzero...
1. Cytochrome alpha 3 in whole-cell suspensions of the fission yeast Schizosaccharomyces pombe reacted in the reduced form with CO to give a photodissociable CO complex with absorption maxima at 429, 543 and 591 nm in CO-liganded reduced-minus-reduced difference spectra. 2. Other CO-bound haemoproteins, cytochromes P-420 and P-450, were not photodissociated under the conditions employed. 3. Measurements of the rates of reassociation of CO with cytochrome alpha 3 after flash photolysis over the temperature range from −101 to −109 degrees C gave a value for Eact. of 28.6 kJ/mol. 4. Between −94 and −106 degrees C, O2 reacted with cytochrome oxidase in intact cells to give an oxygenated intermediate (compound A). 5. At −70 degrees C compound A was converted into a second spectrally distinct intermediate (compound B). 6. Electron transport, indicated by the oxidation of cytochromes alpha + alpha 3 and cytochrome c, did not occur until the temperature was raised to −50 degrees C. 7. At ...http://www.biochemj.org/content/184/3/555
Mitochondrial adenosine triphosphatase of the fission yeast Schizosaccharomyces pombe 972h-. Changes in inhibitor sensitivities...
1. We used 11 different inhibitors of energy conservation as inhibitors of ATPase (adenosine triphosphatase) in extracts of Schizosaccharomyces pombe obtained from cells at different stages of the cell cycle. 2. All the inhibitors showed cell-cycle-dependent variations in their I50 values (microng of inhibitor/mg of protein giving 50% inhibition of inhibitor-sensitive ATPase at pH 8.6). 3. From the sensitivity profiles through the cell cycle it was concluded that: (a) oligomycin, venturicidin, triethyltin sulphate and dibutylchloromethyltin chloride all act at closely associated site(s); (b) NN'-dicyclohexylcarbodi-imide and leucinostatin both act at a similar site, which is, however, distinct from that at which other inhibitors of the membrane factor (Fo) act. 4. The variations in I50 values for efrapeptin closely followed changes in specific activity of ATPase, as would be expected for an inhibitor acting at catalytic sites; these fluctuations were different from those for aurovertin, ...http://www.biochemj.org/content/162/3/581
rad22 - DNA repair and recombination protein rad22 - Schizosaccharomyces pombe (strain 972 / ATCC 24843) (Fission yeast) -...
Active in the repair of DNA damage and in mating-type switching. Probably involved in the repair of DNA double-strands breaks. Has a role in promoting S phase completion.http://www.uniprot.org/uniprot/P36592
SPAC227.10 - Probable prefoldin subunit 2 - Schizosaccharomyces pombe (strain 972 / ATCC 24843) (Fission yeast) - SPAC227.10...
Binds specifically to cytosolic chaperonin (c-CPN) and transfers target proteins to it. Binds to nascent polypeptide chain and promotes folding in an environment in which there are many competing pathways for nonnative proteins (By similarity).http://www.uniprot.org/uniprot/Q9UTC9
Schizosaccharomyces Genomes Project | Broad Institute
Project Information The Schizosaccharomyces Comparative Genome Project project is part of the Broad Institute Fungal Genome Initiative. Its goal was to sequence, annotate and analyze the genomes and transcriptomes of Schizosaccharomyces japonicus, Schizosaccharomyces octosporus and Schizosaccharomyces cryophilus. The primary collaborator for this project was Nick Rhind at the University of Massachusetts, Worcester, Medical School.https://www.broadinstitute.org/scientific-community/science/projects/fungal-genome-initiative/schizosaccharomyces-genomes-project?component=%24DirectLink&service=direct&sp=SSJ5
The Identification of cDNAs That Affect the Mitosis-to-Interphase Transition in Schizosaccharomyces pombe, Including sbp1,...
Yeast strains and cell culture: Schizosaccharomyces pombe strains used were a haploid strain (h− leu1-32 ura4-D18 ade6-m216), a diploid strain (h−/h+ leu1-32/leu1-32 ura4-D18/ura4-D18 ade6-m210/ade6-m216), and a mutant haploid strain pim1-d1ts (h− leu1-32 ura4-D18 pim1-d1ts; Sazer and Nurse 1994), all of which are derived from strain 972 (Leupold 1970). Cell culture conditions, media composition, and genetic analyses have been described previously (Morenoet al. 1991).. nmt1 promoter regulation: Gene expression under the control of the nmt1 promoter (Maundrell 1990) in pREP3X or pREP41X (Forsburg 1993) was repressed by the inclusion of 5 μg/ml thiamine in the Edinburgh Minimal Media (EMM; Morenoet al. 1991). To derepress expression, cells were washed three times with thiamine-free EMM and grown in fresh thiamine-free EMM.. cDNA library screen and DNA manipulations: An S. pombe cDNA library (a gift from Bruce Edgar and Chris Norbury) in the pREP3X vector (Forsburg 1993) was transformed into ...http://www.genetics.org/content/148/2/645
Generation and analysis of a barcode-tagged insertion mutant library in the fission yeast Schizosaccharomyces pombe | BMC...
In this study, we created a fission yeast insertion mutant library in which all mutants were tagged with unique barcode sequences and stored as two readily available selection platforms. The 384-well mutant arrays allow genetic screens on individual mutants and can be extended to genetic approaches such as synthetic genetic array (SGA) [45, 46]. These mutant arrays have been used to identify mutants with four distinct phenotypes (Table 2) as well as strains that are hyper-sensitive to cancer chemotherapeutics camptothecin and bleomycin (Hale and Runge, unpublished data). In addition to 384-well mutant arrays, mutant pools of 1800 mutants are available for parallel analysis.. The insertion mutagenesis used in this study relied on random non-homologous recombination, where a vast majority of transformants have unstable, circularized vector DNA and only a small portion have stable insertions. To facilitate the collection of stable insertion mutants, we included low-dose 5-FOA in our initial ...https://bmcgenomics.biomedcentral.com/articles/10.1186/1471-2164-13-161
The Fission Yeast Schizosaccharomyces pombe Has Two Importin-α Proteins, Imp1p and Cut15p, Which Have Common and Unique...
NUCLEOCYTOPLASMIC transport is a process specific to eukaryotes in which the chromosomes are physically separated from the cytoplasm by the nuclear envelope (NE). In all eukaryotes, the receptor-mediated transport of nuclear proteins across the NE from their site of synthesis in the cytoplasm is essential for all nuclear processes. The precise tissue-specific and temporal regulation of nuclear protein import is also critical in the regulation of cell cycle progression and in developmental and signal transduction pathways (reviewed in Kaffman and O'Shea 1999).. Proteins are targeted to the nucleus by an NLS (nuclear localization signal). There are two types of classical NLSs, both of which must bind to an importin-α adaptor for transport to the nucleus: the mono-partite NLS that consists of 4 or more basic amino acids preceded by a helix-breaking residue and the bipartite NLS that has two short stretches of basic amino acids separated by a 9- to 12-amino-acid spacer (reviewed in Izaurralde and ...http://www.genetics.org/content/171/1/7
Tying the knot: linking cytokinesis to the nuclear cycle | Journal of Cell Science
For the survival of both the parent and the progeny, it is imperative that the process of their physical division (cytokinesis) be precisely coordinated with progression through the mitotic cell cycle. Recent studies in the budding yeast Saccharomyces cerevisiae and the fission yeast Schizosaccharomyces pombe are beginning to unravel the nature of the links between cytokinesis and the nuclear division cycle. The cyclin-dependent kinases and a novel surveillance mechanism that monitors cytokinesis and/or morphogenesis appear to play important regulatory roles in forging these links. It is becoming increasingly clear that the inactivation of the mitosis-promoting cyclin-dependent kinase, which marks the completion of the nuclear division cycle, is essential for actomyosin ring constriction and division septum assembly in both yeasts. Additionally, the spindle pole bodies are emerging as important transient locale for proteins that might play a key role in coupling the completion of mitosis to ...http://jcs.biologists.org/content/113/9/1503
"Sid2p, a spindle pole body kinase that regulates the onset of cytokine" by Cynthia A. Sparks, Mary K. Morphew et al.
The fission yeast Schizosaccharomyces pombe divides by medial fission through the use of an actomyosin contractile ring. Precisely at the end of anaphase, the ring begins to constrict and the septum forms. Proper coordination of cell division with mitosis is crucial to ensure proper segregation of chromosomes to daughter cells. The Sid2p kinase is one of several proteins that function as part of a novel signaling pathway required for initiation of medial ring constriction and septation. Here, we show that Sid2p is a component of the spindle pole body at all stages of the cell cycle and localizes transiently to the cell division site during medial ring constriction and septation. A medial ring and an intact microtubule cytoskeleton are required for the localization of Sid2p to the division site. We have established an in vitro assay for measuring Sid2p kinase activity, and found that Sid2p kinase activity peaks during medial ring constriction and septation. Both Sid2p localization to the ...https://escholarship.umassmed.edu/oapubs/933/
Regulation of Schizosaccharomyces pombe Atf1 protein levels by Sty1-mediated...
The Atf1 transcription factor plays a vital role in the ability of Schizosaccharomyces pombe cells to respond to various stress conditions. It regulates the expression of many genes in a stress-dependent manner, and its function is dependent upon the stress-activated MAPK, Sty1/Spc1. Moreover, Atf1 is directly phosphorylated by Sty1. Here we have investigated the role of such phosphorylation. Atf1 protein accumulates following stress, and this accumulation is lost in a strain defective in the Sty1 signaling pathway. In addition, accumulation of a mutant Atf1 protein that can no longer be phosphorylated is lost. Measurement of the half-life of Atf1 demonstrates that changes in Atf1 stability are responsible for this accumulation. Atf1 stability is also regulated by its heterodimeric partner, Pcr1. Similarly, Pcr1 levels are regulated by Atf1. Thus multiple pathways exist that ensure that Atf1 levels are appropriately regulated. Phosphorylation of Atf1 is important for cells to mount a robust ...http://christie.openrepository.com/christie/handle/10541/71277
Conditional inactivation of replication proteins in fission yeast using hormone-binding domains. - Oxford Neuroscience
The fission yeast Schizosaccharomyces pombe is a useful model for analysing DNA replication as genetic methods to allow conditional inactivation of relevant proteins can provide important information about S-phase execution. A number of strategies are available to allow regulation of protein level or activity but there are disadvantages specific to each method and this may have limitations for particular proteins or experiments. We have investigated the utility of the inducible hormone-binding domain (HBD) system, which has been described in other organisms but little used in fission yeast, for the creation of conditional-lethal replication mutants. In this method, proteins are tagged with HBD and can be regulated with β-estradiol. In this article, we describe the application of this method in fission yeast, specifically with regard to analysis of the function of GINS, an essential component of the eukaryotic replicative helicase, the CMG complex.https://www.neuroscience.ox.ac.uk/publications/325474
Degradation of HMG-CoA reductase-induced membranes in the fission yeast, Schizosaccharomyces pombe. | JCB
Elevated levels of certain membrane proteins, including the sterol biosynthetic enzyme HMG-CoA reductase, induce proliferation of the endoplasmic reticulum. When the amounts of these proteins return to basal levels, the proliferated membranes are degraded, but the molecular details of this degradation remain unknown. We have examined the degradation of HMG-CoA reductase-induced membranes in the fission yeast, Schizosaccharomyces pombe. In this yeast, increased levels of the Saccharomyces cerevisiae HMG-CoA reductase isozyme encoded by HMG1 induced several types of membranes, including karmellae, which formed a cap of stacked membranes that partially surrounded the nucleus. When expression of HMG1 was repressed, the karmellae detached from the nucleus and formed concentric, multilayered membrane whorls that were then degraded. During the degradation process, CDCFDA-stained compartments distinct from preexisting vacuoles formed within the interior of the whorls. In addition to these ...http://jcb.rupress.org/content/131/1/81
Fission yeast Pds5 is required for accurate chromosome segregation and for survival after DNA damage or metaphase arrest. -...
Sister chromatid cohesion, which is established during the S phase of the eukaryotic cell cycle and persists until the onset of anaphase, is essential for the maintenance of genomic integrity. Cohesion requires the multi-protein complex cohesin, as well as a number of accessory proteins including Pds5/BIMD/Spo76. In the budding yeast Saccharomyces cerevisiae Pds5 is an essential protein that localises to chromosomes in a cohesin-dependent manner. Here we describe the characterisation in the fission yeast Schizosaccharomyces pombe of pds5(+), a novel, non-essential orthologue of S. cerevisiae PDS5. The S. pombe Pds5 protein was localised to punctate nuclear foci in a manner that was dependent on the Rad21 cohesin component. This, together with additional genetic evidence, points towards an involvement of S. pombe Pds5 in sister chromatid cohesion. S. pombe pds5 mutants were hypersensitive to DNA damage and to mitotic metaphase delay, but this sensitivity was apparently not due to precocious ...https://www.neuroscience.ox.ac.uk/publications/2768
A mutated dph3 gene causes sensitivity of Schizosaccharomyces pombe cells to cytotoxic agents (pdf) | Paperity
Paperity: the 1st multidisciplinary aggregator of Open Access journals & papers. Free fulltext PDF articles from hundreds of disciplines, all in one placehttp://paperity.org/p/79890276/a-mutated-dph3-gene-causes-sensitivity-of-schizosaccharomyces-pombe-cells-to-cytotoxic
Correct Regulation of the Septation Initiation Network in Schizosaccharomyces pombe Requires the Activities of par1 and par2 |...
Cytokinesis is a crucial event in cell division. It needs to occur at the right time, in the right place, and only once during a single cell cycle. As such, it must be highly regulated. We have presented evidence showing that par1- and par2-directed PP2A activity is required for proper functioning of the S. pombe SIN pathway. This activity is required at the level of the Ras-like GTPase, Spg1p, whose function is situated at the beginning of this signaling pathway. It appears that the role played by par1 and par2 is to negatively regulate SIN, thereby ensuring that multiple rounds of septation do not occur.. The suppression of spg1-106 by par1Δ is specific to the restoration of the SIN pathway function: As a strain with a loss-of-function ts allele of spg1, spg1-106 cells displayed a typical sid phenotype when shifted to 36°, their restrictive temperature. Deleting par1 in this strain strongly suppressed both the morphological and growth defects of spg1-106 cells. When par1Δ spg1-106 double ...http://www.genetics.org/content/158/4/1413
Suppression of the Schizosaccharomyces pombe cut12.1 cell-cycle defect by mutations in cdc25 and genes involved in...
Cdc25 phosphatase primes entry to mitosis by removing the inhibitory phosphate that is transferred to mitosis promoting factor (MPF) by Wee1 related kinases. A positive feedback loop then boosts Cdc25 and represses Wee1 activities to drive full-scale MPF activation and commitment to mitosis. Dominant mutations in the Schizosaccharomyces pombe spindle pole body (SPB) component Cut12 enable cdc25.22 mutants to overcome a G2 arrest at 36 degrees and enter mitosis. The recessive temperature-sensitive cut12.1 mutation results in the formation of monopolar spindles in which the spindle pole marker Sad1 is enriched on the nonfunctional SPB at 36 degrees . We identified mutations at five loci that suppressed the lethality of the recessive cut12.1 mutation at 36 degrees and conferred lethality at 20 degrees . Three of the five mutations led to the formation of monopolar spindles at restrictive temperatures, affected cell size at commitment to mitosis, and generated multiple Sad1 foci at nuclear ...http://christie.openrepository.com/christie/handle/10541/71916
Using the DHFR heat-inducible degron for protein inactivation in Schizosaccharomyces pombe. - Oxford Neuroscience
Inactivating a specific protein in vivo can yield important information about its function. One strategy previously developed in Saccharomyces cerevisiae by the Varshavsky group involves fusing a degron, derived from mouse dihydrofolate reductase, to the N-terminus of the target protein, which thereby confers temperature-sensitive degradation at the restrictive temperature. We describe here the application of this technique in the fission yeast, Schizosaccharomyces pombe.https://www.neuroscience.ox.ac.uk/publications/209500
Fission yeast Prp4p kinase regulates pre‐mRNA splicing by phosphorylating a non‐SR‐splicing factor | EMBO Reports
The recognition and removal of introns in eukaryotes is mediated by a highly complex and dynamic machinery called the spliceosome (Will and Lührmann, 1997). The basic RNA and protein components of this machinery have been well conserved throughout evolution; however, we found distinct differences when we compared the splicing factors of fission yeast Schizosaccharomyces pombe with those of Homo sapiens and the budding yeast Saccharomyces cerevisiae (Käufer and Potashkin, 2000).. The modification of some of the proteins by phosphorylation appears to play an important role in the splicing process. It has been shown that kinase activity in mammals is essential for pre‐mRNA splicing in vitro (Tazi et al., 1993; Mermoud et al., 1994). In mammals, several protein kinases have been identified that specifically phosphorylate the RS domain of the SR‐splicing factors in vitro (Gui et al., 1994; Colwill et al., 1996; Rossi et al., 1996). SR proteins consisting of one or two RNA‐binding domains ...http://embor.embopress.org/content/2/1/35
British Library EThOS: DNA replication in Schizosaccharomyces pombe
The primary aim of this thesis was to isolate temperature sensitive genetic mutants of Schizosaccharomyces pombe which are defective in DNA replication. 30 presumptive. (DNA) mutants were isolated after nitrosoguanidine mutagenesis on the basis of their having a defect in the S phase period of the cell cycle. The mutations are all recessive and were allocated to 9 unlinked nuclear genes (cde 10, 17, 18, 19, 20, 21, 22, 23, 24). Physiological characterization revealed that (1) Mutants of cdc 10, 20, 22 are probably defective in the initiation of S phase, (2) mutants of cdc 21, 23 are defective in the process of DNA synthesis, (3) mutants of cdc 18, 19 are not defective in bulk DNA synthesis and their to functions are completed prior to or independently of S phase at the permissive temperature, and (4) mutants of cdc 17, 2.4 are not defective in bulk DNA synthesis but their is functions cannot be completed at the permissive temperature if DNA synthesis is inhibited by hydro urea. cdc 17-K42 ...http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.659887
Fission yeast TORC1 prevents eIF2α phosphorylation in response to nitrogen and amino acids via Gcn2 kinase | Journal of Cell...
In response to diverse environmental stresses, the phosphorylation of eukaryotic initiation factor-2 (eIF2α) induces a programme of gene expression that mitigates cellular injury. Several protein kinases phosphorylate eIF2α at serine 51, leading to the inhibition of eIF2B activity, that in turn causes a reduction in general protein synthesis and an enhancement of the translation of specific mRNAs encoding for proteins that remediate the stress. In mammalian cells, four eIF2 kinases (HRI, GCN2, PEK/Perk and PKR) inhibit translation initiation through the phosphorylation of eIF2α in response to different types of cellular stress (Dever, 2002; Proud, 2005). In Saccharomyces cerevisiae, Gcn2p is activated upon nutrient limitation (amino acids, purine and glucose), but also by high concentrations of sodium, rapamycin and methyl methanesulfonate (Cherkasova and Hinnebusch, 2003; Narasimhan et al., 2004; Valenzuela et al., 2001; Yang et al., 2000). In the fission yeast Schizosaccharomyces pombe, ...http://jcs.biologists.org/content/125/24/5955
The Schizosaccharomyces pombe HIRA-like protein hip1 is required for the periodic expression of histone genes and contributes...
The Schizosaccharomyces pombe HIRA-like protein hip1 is required for the periodic expression of histone genes and contributes to the function of complex ...http://eprint.ncl.ac.uk/pub_details2.aspx?pub_id=69512
alg9 Gene, alg9 Transcript, alg9 Protein, and alg9 Antibody | Schizosaccharomyces pombe | RefGene
Schizosaccharomyces pombe. alg9 gene, alg9 microRNA and miRNA, alg9 transcript, and alg9 antibody; mannosyltransferase complex subunit Alg9 (predicted); - alg9 RefGene Summary.http://refgene.com/gene/2542331
A geographically diverse collection of schizosaccharomyces pombe isolates shows limited phenotypic variation but extensive...
Brown, William R.A. and Litti, Gianni and Rosa, Carlos and James, Steve and Roberts, Ian and Robert, Vincent and Jolly, Neil and Tang, Wen and Baumann, Peter and Green, Carter and Schlegel, Kristina and Young, Jonathan and Hirchaud, Fabienne and Leek, Spencer and Thomas, Geraint and Blomberg, Anders and Warringer, Jonas (2011) A geographically diverse collection of schizosaccharomyces pombe isolates shows limited phenotypic variation but extensive karyotypic diversity. G3, 1 (7). pp. 615-626. ISSN 2160-1836 ...http://eprints.nottingham.ac.uk/2978/
A microPIXE investigation of the interaction of cells of Schizosaccharomyces pombe with the culture medium - Surrey Research...
Rombouts, PMM, Gomez-Morilla, I, Grime, GW, Webb, RP, Cuenca, L, Rodriguez, R, Browton, M, Wardell, N, Underwood, B, Kirkby, NF et al and Kirkby, KJ. (2007) A microPIXE investigation of the interaction of cells of Schizosaccharomyces pombe with the culture medium In: 10th International Conference on Nuclear Microprobe Technology and Applications held in Conjunction with the 2nd International Workshop on Proton Beam Writing, 2006-07-09 - 2006-07-14, Singapore, SINGAPORE. Full text not available from this repository ...http://epubs.surrey.ac.uk/829396/