TY - JOUR. T1 - Differential mRNA expression of prostaglandin receptor subtypes in macrophage activation. AU - Hubbard, Neil. AU - Lee, S. H.. AU - Lim, D.. AU - Erickson, Kent L. PY - 2001. Y1 - 2001. N2 - Assessing the regulation of macrophage receptors for prostaglandin (PGE2) is essential to understanding the control which that potent lipid mediator has in modulating macrophage activities. The purpose of this study was to assess the differential mRNA expression of PGE2 receptor subtypes (EP) during macrophage exposure to activating and transducing agents. RAW 264.7 macrophages constitutively expressed mRNA for EP2, EP3 and EP4 receptor subtypes. Messenger RNA for EP4 was expressed at a much higher level when compared to EP2 in unstimulated macrophages as assessed by kinetic quantitative RT-PCR. When macrophages were stimulated with LPS, EP2 mRNA levels were 12-fold higher when compared to unstimulated macrophages, while EP4 mRNA remained unchanged. Conversely, mRNA levels of both EP2 and EP4 ...
The substantial amount of RNA required for expression analysis is a limiting factor for the cDNA microarray technology in a number of potentially important applications. Two main approaches, signal amplification and global mRNA amplification, have been developed to overcome this obstacle. Signal amplification, such as dendrimer technology [1] and tyramide signal amplification (TSA) [2] aim to increase the fluorescent signal emitted per mRNA molecule. Global mRNA amplification has the purpose of increasing the number of available transcript equivalents for sufficient labeling from a limited starting amount. In current implementations, mRNA amplification techniques require less RNA than those based on signal amplification.. Van Gelder et al. [3] devised a multistep strategy to amplify mRNA from limited quantities of cDNA in studies of gene expression. Their method is commonly referred to in the literature as the Eberwine method. The general steps involve reverse transcription of mRNA with an oligo ...
TY - JOUR. T1 - BCL2 protein expression parallels its mRNA level in normal and malignant B cells. AU - Shen, Yulei. AU - Iqbal, Javeed. AU - Huang, James Z.. AU - Zhou, Guimei. AU - Chan, Wing C.. PY - 2004/11/1. Y1 - 2004/11/1. N2 - The regulation of B-cell lymphoma 2 (BCL2) protein expression in germinal center (GC) B cells has been controversial. Previous reports have indicated post-transcriptional regulation plays a dominant role. However, a number of recent studies contradicted these reports. Using real-time polymerase chain reaction (PCR) and Standardized Reverse Transcriptase-PCR (StaRT-PCR), we measured the level of mRNA expression in GC, mantle zone (MNZ), and marginal zone (MGZ) cells from laser capture microdissection. Both quantitative RT-PCR measurements of microdissected GC cells from tonsils showed that GC cells had low expression of BCL2 transcripts commensurate with the low protein expression level. These results are in agreement with microarray studies on fluorescence-activated ...
Figure 3 Thermogenic program and β3-adrenergic receptor signaling is mediated by membrane-initiated ERα signaling. A: Immunoblot analysis of UCP1 levels in BAT and pWAT of ERα+/+, ERα−/−, WT, and KRRki/ki mice. Representative immunoblots and quantification are shown (n = 5-8 per group). #P , 0.01. B: qRT-PCR analysis of genes consistent with beige adipocytes in adipose tissues of ERα+/+, ERα−/−, WT, and KRRki/ki mice (n = 6-8). Relative mRNA expression levels are normalized to gapdh. *P , 0.05, #P , 0.01. C: Hematoxylin and eosin staining of pWAT of ERα+/+, ERα−/−, WT, and KRRki/ki mice. Scale bar indicates 100 µm. The graph depicts the quantification of mean cell area (n = 4). #P , 0.01. D: qRT-PCR analysis of genes consistent with beige adipocytes in NIH 3T3-L1 preadipocytes treated with vehicle (control), 100 nmol/L E2, or 2 µmol/L rosiglitazone for 72 h. Relative mRNA expression levels are normalized to gapdh. Data depict the results from three independent experiments. ...
Non-small cell lung tumor (NSCLC) may be the most common tumor as well as the leading reason behind death from tumor worldwide. in comparison to people that have low mRNA amounts (20.three months vs 34.three months, respectively; Log Rank Check, p?=?0.016), when contemplating all NSCLC levels which difference is buy 958772-66-2 a great deal larger when contemplating only sufferers with stage IV (15.9 months vs 31.three months, respectively; Log Rank Check, p?=?0.036). Furthermore, circulating Ang-2 mRNA amounts independently determine general survival, as well as the concordance (c) index evaluation showed that this is of the nomogram which has information relating to tumor stage, sufferers smoking position and circulating Ang-2 mRNA amounts present an elevated capacity to anticipate overall success in NSCLC sufferers (c-index 0.798). These outcomes claim that this nomogram could serve as a distinctive and practical device to determine prognosis in NSCLC, not really counting on the option of ...
RNA expression patterns of cancer-adjacent breast tissue could be to gauge future survival outcomes for women with estrogen receptor-positive breast cancer
Gene expression differs among individuals and populations and is thought to be a major determinant of phenotypic variation. Although variation and genetic loci responsible for RNA expression levels have been analysed extensively in human populations, our knowledge is limited regarding the differences in human protein abundance and the genetic basis for this difference. Variation in messenger RNA expression is not a perfect surrogate for protein expression because the latter is influenced by an array of post-transcriptional regulatory mechanisms, and, empirically, the correlation between protein and mRNA levels is generally modest. Here we used isobaric tag-based quantitative mass spectrometry to determine relative protein levels of 5,953 genes in lymphoblastoid cell lines from 95 diverse individuals genotyped in the HapMap Project. We found that protein levels are heritable molecular phenotypes that exhibit considerable variation between individuals, populations and sexes. Levels of specific sets of
Figure 3. Western blots of Cos7 cell extracts stained with CCM2 antibody (A and B). Cells were transfected with an expression vector encoding a full-length CCM2-GFP fusion protein ( ). Untransfected cells served as controls ( ). A, In transfected cells, the CCM2 antibody detects an 82-kDa protein (expected size for the fusion protein). No staining is observed in untransfected controls. B, Peptide competition eliminates staining, demonstrating the specificity of this antibody for the CCM2 protein. C, Multitissue Western blot reveals CCM2 protein expression in the brain, heart, lung, and kidney. - CCM2 expression parallels that of CCM1.
Principal Investigator:FUKUDA Takeshi, Project Period (FY):1991 - 1993, Research Category:Grant-in-Aid for General Scientific Research (C), Research Field:Respiratory organ internal medicine
gene, would facilitate examination of the role of this gene in the inheritance of human obesity. Northern blot analysis revealed that OB RNA is present at a high level in adipose tissue but at much lower levels in placenta and heart. OB RNA is undetectable in a wide range of other tissues. Comparative mapping of mouse and human DNA indicated that the ob gene is located within a region of mouse chromosome 6 that is homologous to a portion of human chromosome 7q. We mapped the human ...
Individual fractions of polysomes were isolated from yeast. Pulse labeling experiments in vivo show constant specific activity of messenger RNA in each polysome peak; this suggests a uniform density of ribosomes per unit length of messenger RNA. In the cell-free incorporating system, the amount of peptide per ribosome unit increased with the size of polysome.. ...
酵母细胞通过提高葡萄糖合成酶和糖酵解作用维持能量代谢的平衡。在人体细胞中我们也发现了同样的代谢调节途径,主要通过eIF3e蛋白与代谢相关的mRNA结合并促进这些与能量代谢相关的蛋白表达来维持代谢的平衡。在肿瘤形成的过程中,这种通过eIF3d和eIF3e调控的能量代谢机制是被打破的,因此该研究成果将有助于科学家通过靶向代谢通路研发治愈肿瘤的药物。该研究利用转录组学、蛋白组学和代谢组学的方法发现缺少了eIF3e和eIF3d的裂殖酵母细胞无法合成线粒体电子传递链相关的蛋白,从而导致呼吸功能阻断,内源性的氧化应激压力和细胞老化产生。 课题组硕士研究生苏丹为共同第一作者 。 Dieter A. ...
Shinde and Klein are not yet sure whether GSK-3s effect on RNA splicing explains its role in mood disorders. The effect of GSK-3 on messenger RNA in neuronal cells, with or without lithium, would need to be examined to determine this. The study underlines how investigations into the basic biological function of a drug target can lead in unexpected directions. "[The GSK-3 phosphoproteome] is a really large data set," Shinde said. "Its a resource for the field." "The relevance to leukemia could be direct and something worthy of immediate study," Klein said. "The role in psychiatric disorders is a major interest of the work, but the impact would be down the road, not immediate." ...
Time-dependent induction of connected cells by overexpression of cdc5ΔN. (A) Strain KLY1083 expressing three copies of GAL1-EGFP-cdc5ΔN homogeneously induced
Alternative splicing (AS) is a post-transcriptional regulatory mechanism for gene expression regulation. Splicing decisions are affected by the combinatorial behavior of different splicing factors that bind to multiple binding sites in exons and introns.
... (mRNA) is a molecule of RNA encoding a chemical "blueprint" for a protein product. mRNA is transcribed from a DNA template, and carries coding information to the sites of protein synthesis: the ribosomes. Here, the nucleic acid polymer is translated into a polymer of amino acids: a protein. In mRNA as in DNA, genetic information is encoded in the sequence of nucleotides arranged into codons consisting of three bases each. Each codon encodes for a specific amino acid, except the stop codons that terminate protein synthesis. This process requires two other types of RNA: transfer RNA (tRNA) mediates recognition of the codon and provides the corresponding amino acid, while ribosomal RNA (rRNA) is the central component of the ribosomes protein manufacturing machinery.. ...
Background: Current methodology of gene expression analysis limits the possibilities of comparison between cells/tissues of organs in which cell size and/or number changes as a consequence of the study (e.g. starvation). A method relating the abundance of specific mRNA copies per cell may allow direct comparison or different organs and/or changing physiological conditions. Methods: With a number of selected genes, we analysed the relationship of the number of bases and the fluorescence recorded at a present level using cDNA standards. A lineal relationship was found between the final number of bases and the length of the transcript. The constants of this equation and those of the relationship between fluorescence and number of bases in cDNA were determined and a general equation linking the length of the transcript and the initial number of copies of mRNA was deduced for a given pre-established fluorescence setting. This allowed the calculation of the concentration of the corresponding mRNAs per ...
I would like to know what amounts of protein are necessary for pregnant women? I have been eating the Paleo Diet since you introduced me to it. This is my first
Im prepping for surgery right now with a liquid diet. What would you say is the minimum grams of protein to get a day? My doctor wants 100g but Ive been lucky to get 60g. Is this going to hurt me down the road?Thanks!
YV Subrahmanyam, S Yamaga, Y Prashar, HH Lee, NP Hoe, Y Kluger, M Gerstein, JD Goguen, PE Newburger, SM Weissman (2001). Blood 97: 2457-68 ...
Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse monocytic and immortalized Mm-1 cells derived from spontaneously differentiated, mouse myeloblastic M1 cells. Although these subline cells retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better understand the molecular mechanisms of these levels of leukemogenicity, we investigated putative leukemogenesis-associated genes or oncogenes involved in the maintenance of growth, especially in vivo, by means of differential mRNA display. We isolated a fragment clone (15T01) from Mm-P cells. The mRNA probed with 15T01 was expressed at high levels in leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2 cells. The gene corresponding to 15T01, named TRA1, was isolated from an Mm-P cDNA library. The longest open reading frame of the TRA1 clone predicts ...
Mouse monocytic Mm-A, Mm-P, Mm-S1, and Mm-S2 cells are sublines of mouse monocytic and immortalized Mm-1 cells derived from spontaneously differentiated, mouse myeloblastic M1 cells. Although these subline cells retain their monocytic characteristics in vitro, Mm-A and Mm-P cells are highly leukemogenic to syngeneic SL mice and athymic nude mice, whereas Mm-S1 and Mm-S2 cells are not or are only slightly leukemogenic. To better understand the molecular mechanisms of these levels of leukemogenicity, we investigated putative leukemogenesis-associated genes or oncogenes involved in the maintenance of growth, especially in vivo, by means of differential mRNA display. We isolated a fragment clone (15T01) from Mm-P cells. The mRNA probed with 15T01 was expressed at high levels in leukemogenic Mm-P and Mm-A cells but not in nonleukemogenic Mm-S1 and Mm-S2 cells. The gene corresponding to 15T01, named TRA1, was isolated from an Mm-P cDNA library. The longest open reading frame of the TRA1 clone predicts ...
Cancer cells. (A) Relative expression levels of Nox1, 2, 3, 4, and 5 mRNAs in A549 cells were determined by real-time RT-PCR and are presented as mean delta Ct
J:60127 Pascolo S, Tsoukatou D, Mamalaki C, Identification of thymus specific and developmentally regulated genes by an improved version of the mRNA differential display technique. Dev Immunol. 1999;7(1):1-7 ...
Comments, concepts and statistics about Systematic discovery of structural elements governing stability of mammalian messenger RNAs..
The technology we have available to us today in the lab is both a boon and a bafflement. Example: The screens we have for RNA expression in cells is so sensitive we can see tiny changes in RNA expression levels in healthy/diseased/drug treated/etc cells. YAY! More information! More observations! More new ideas for research!… Except,…. ...
DS was born at 36 weeks by EMCS, following a failed induction attempt. There were complications in the pregnancy that meant he was monitored closely
DNA is the informational basis from which living cells derive instructions for synthesizing proteins. Many of the resulting proteins are enzymes that catalyze biochemical reactions from which the cell derives energy or generates other molecules essential to its health and safety. The process normally occurs when the sequence of nucleotides in DNA is transcribed into a complementary, single strand of nucleotides known as messenger RNA, or mRNA. The mRNA provides the instructions by which other components in the cell synthesize proteins. Because not all genes are transcribed (or expressed) but all genes that are transcribed do so through mRNA, the presence of mRNA is an indicator that a gene from the cells DNA has been expressed. The DNA from which the mRNA is obtained is sometimes interspersed with oligonucleotide spacers that do not appear in the final mRNA. Because mRNA is used for the cells molecular machinery to generate the protein, the sequence of DNA (or gene) that corresponds to a ...
overexpressing GPX1 in endothelial cells is able to change the basal mRNA and protein BAX levels without affecting those of TP53 and BCL2 (useful to antiatherogenic therapies which use antioxidants with the aim of protecting the vascular wall against ...
Question 1 (1 point) Which of the following is a true statement about genes?Question 1 options: A) Genes are structures within chromosomes of each cell that contain deoxyribonucleic acid B) Genes are messenger ribonucleic acid (mRNA) C) Genes are cells that determine gender during fertilization D) Genes are antibodies that promote disease SaveQuestion 2 (1 point) Which of the following is the major cause of death from cancer?Question 2 options: A) Infection B) Hemorrhage C) Pain D) Metastasis SaveQuestion 3 (1 point) Which of the following statements is true with respect to the stages of cancer?Question 3 options: A) Stage 1 represents a poor prognosis B) Localized cancer is a stage 4 C) Benign tumors are stage 2 D) &
CureVac is the first company to successfully harness messenger RNA (mRNA) for medical purposes. The principle is promising: use natural mRNA as a data carrier to instruct the human body to produce its own proteins to fight a wide range of diseases. ...
Study Flashcards On FA page 199 G-protein linked 2nd messengers, major functions plus major grouping at Cram.com. Quickly memorize the terms, phrases and much more. Cram.com makes it easy to get the grade you want!
Last weekend I had the opportunity to learn from one of the most intelligent fitness minds in the industry. Aside from learning an abundant amount of
Total RNAs are available in human, mouse, rat, and cow. In addition to normal tissues, a large number of human fetal, tumor and cell line Total RNAs are available
Total RNAs are available in human, mouse, rat, and cow. In addition to normal tissues, a large number of human fetal, tumor and cell line Total RNAs are available
With all of thre recent talk of the noes, I wanted to post pics of my mono noe worn messenger style with the long strap (purchased separately for...
Translate Bio, Inc. (TBIO), a clinical-stage messenger RNA (mRNA) therapeutics company developing a new class of potentially transformative medicines to treat diseases caused by protein or gene dysfunction, today announced the pricing of its underwritten public offering of 9,000,000 shares of its common
WE have seen how the ordinary glands of the body produce their secretions and how the latter pass down particular ducts to the sites where they act. We have now
டி.என்.ஏ யில் இருந்து ஆர்.என்.ஏ யாக மாற்றப்படும் செயல்முறை. இம்மாற்றத்தின் போது மரபு ஈரிழையின் (டி.என்.எ ) 5 (முனை அல்லது தொடர்) ப்ரீம் இருந்து 3 (முனை அல்லது தொடர்) ப்ரீம் நோக்கி ஆர்.என்.ஏ நகலாக்கப்படும். இவ்வினையின் பொழுது ஆர்.என்.ஏ பாலிமரசு ௨ (II) என்ற நொதி டி.என்.ஏ யிலிருந்து செய்திகாவும் (messenger) ஆர்.என்.ஏ (எம்.ஆர்.என்.ஏ) உருவாதலில் முக்கிய பங்காற்றுகின்றது. இதுவே ஆர்.என்.ஏ. படியெடுப்பு ...
The TV MegaSites As The World Turns Site is a large fan page with information, links, daily summaries, transcripts, and more
The TV MegaSites As The World Turns Site is a large fan page with information, links, daily summaries, transcripts, and more
Promoter Methylation and Relative mRNA Expression of the p16 Gene in Cervical Cancer in North Indians Cervical cancer;p16 methylation;p16 mRNA expression;North Indian population; Background: Cervical carcinoma is one of the main causes of mortality in women worldwide as well as in India. It occurs as a result of various molecular events that develop from the combined influences of an individuals genetic predisposition and external agents such as smoking and menstrual hygiene, for example. However, infection with human papillomavirus (HPV) is the established major risk factor. The aim of the current study was to investigate p16 CpG island methylation and establish any correlation with mRNA expression in north Indian population. Materials and Methods: We analyzed 196 woman volunteer out of which 98 were cases and 98 healthy controls. For the analysis of methylation pattern, DNA extracted from blood samples was modified with a bisulfate kit and used as template for methylation specific PCR (MSP).
The regulation of mRNA stability has emerged as a critical control step in dynamic gene expression. This process occurs in response to modifications of the cellular environment, including hormonal variations, and regulates the expression of subsets of proteins whose levels need to be rapidly adjusted. Modulation of messenger RNA stability is usually mediated by stabilizing or destabilizing RNA-binding proteins (RNA-BP) that bind to the 3-untranslated region (3UTR) regulatory motifs, such as AU-rich elements (AREs). Destabilizing ARE-binding proteins enhance the decay of their target transcripts by recruiting the mRNA decay machineries. Failure of such mechanisms, in particular misexpression of RNA-BP, has been linked to several human diseases. In the adrenal cortex, the expression and activity of mRNA stability regulatory proteins are still understudied. However, ACTH- or cAMP-elicited changes in the expression/phosphorylation status of the major mRNA-destabilizing protein TIS11b/BRF1 or in the
TY - JOUR. T1 - Myogenic, matrix and growth factor mRNA expression in human skeletal muscle: effect of contraction intensity and feeding. AU - Agergaard, Jakob. AU - Reitelseder, Søren. AU - Pedersen, T.G.. AU - Doessing, Simon. AU - Schjerling, Peter. AU - Langberg, Henning. AU - Miller, Benjamin F. AU - Aagaard, P. AU - Kjær, Michael. AU - Holm, Lars. PY - 2013/3/20. Y1 - 2013/3/20. N2 - INTRODUCTION: We examined short-term (3-hour) and long-term (12-week) training effects after heavy load [HL; 70% 1RM] and light load (LL; 16% 1RM) exercise. METHODS: mRNA expression of genes involved in skeletal muscle remodeling were analyzed and muscle activity (EMG measurements) was measured. RESULTS: Relative muscle activity differed between HL and LL resistance exercise, whereas median power frequency was even, suggesting an equal muscle-fiber-type recruitment distribution. mRNA expression of Myf6, myogenin, and p21 was mostly increased, and myostatin was mostly depressed by HL resistance exercise. No ...
The overall aim of this thesis was to investigate how genome engineering might be used to generate Escherichia coli strains with increased capacities for recombinant protein production. As translation constitute a possible bottleneck in recombinant production processes [Mahalik et. al, 2014] this work focused on evaluating and increasing translational capacities in E. coli. Two methods were used to evaluate translational capacities in E. coli: 1. Two plasmid reporter systems were established to investigate levels of ribosome expression. These plasmids carried red fluorescent protein genes (mCherry), under control of ribosomal promoters. Levels of expression of ribosomal constituents were evaluated by measuring fluorescence from strains carrying reporter plasmids.. 2. Exponential phase growth rates were used to assess translational capacities. Protein synthesis is generally the growth rate limiting factor during exponential phases, and a positive linear correlation between growth rates and ...
This study examined the different molecular forms of CRH in normal and preeclampsia maternal plasma and protease-blocked placental extracts using antibodies to different regions of the CRH precursor, pro-CRH. In the absence of protease inhibitors, chromatographed normal placental extracts contained four peaks of immunoreactivity corresponding to unprocessed approximately 19-kDa pro-CRH, its approximately 8-kDa intermediate metabolite, pro-CRH125-194, its approximately 2.8-kDa midportion fragment, pro-CRH125-151, and 4.75-kDa CRH1-41. However, if protease inhibitors were included in the extraction medium, only pro-CRH and pro-CRH125-194 were found. Pro-CRH processing was more extensive in protease-blocked preeclampsia placentas than in those from normal pregnancy, with three peaks corresponding to pro-CRH, pro-CRH125-194, and mature CRH1-41 peptide found. Using quantitative competitive PCR, the messenger ribonucleic acid levels of CRH precursor in preeclampsia placentas were 1.7-fold higher than ...
The desired end point for the description of a biological system is not the analysis of mRNA transcript levels alone but also the accurate measurement of protein expression levels and their respective activities. Quantitative analysis of global mRNA levels currently is a preferred method for the analysis of the state of cells and tissues (11). Several methods which either provide absolute mRNA abundance (34, 35) or relative mRNA levels in comparative analyses (20, 27) have been described elsewhere. The techniques are fast and exquisitely sensitive and can provide mRNA abundance for potentially any expressed gene. Measured mRNA levels are often implicitly or explicitly extrapolated to indicate the levels of activity of the corresponding protein in the cell. Quantitative analysis of protein expression levels (proteome analysis) is much more time-consuming because proteins are analyzed sequentially one by one and is not general because analyses are limited to the relatively highly expressed ...
In order to better understand cellular responses to viral infection at the transcriptional level, we employed differential display RT-PCR to analyze mRNAs from RD rhabdomyosarcoma cells following infection with a neurovirulent enterovirus 71 (EV71) strain, compared with mRNAs from uninfected cells. Of 250 expressed sequence tags (ESTs) isolated, sequenced, and identified, all were of cellular origin except 1 that was of viral origin. Of these, 156 were individual distinctive clones, comprising 45 mRNAs showing unaltered expression and 111 mRNAs exhibiting upregulation or downregulation. Of the 45 uniformly expressed mRNAs, 14 represented unknown genes. Of the 111 differentially expressed mRNAs, 63 did not match any known genes. Forty-eight of the 111 mRNAs modified by EV71 infection matched known genes, including those encoding components of cell cycle, cytoskeleton, and cell death mediators; protein degradation mediators; mitochondrial-related proteins; components of protein translation and ...
The presence and abundance of 5-HT1A and 5-HT2A receptor mRNAs in post mortem human hippocampus was investigated using a novel quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) technique using cyclophilin mRNA as an internal standard. 5-HT1A and 5-HT2A receptor mRNAs were each co-amplified with varying dilutions of cyclophilin primers, and their abundance expressed as a ratio of cyclophilin mRNA. Using this technique in combination with quantitative autoradiography we have investigated the effect of aging on hippocampal 5-HT1A and 5-HT2A receptor mRNA abundance and binding site densities. There was a significant negative correlation between hippocampal 5-HT1A receptor binding site densities and age and a similar trend for 5-HT1A receptor mRNA abundance. Neither 5-HT2A receptor binding site densities nor mRNA abundance were affected by age. Both 5-HT1A and 5-HT2A receptor binding site densities in individual subjects correlated significantly with abundance of their encoding mRNA. This
In our study, we tried to quantify the relative contribution of transcriptional and post‐transcriptional regulation to mRNA levels. We show that tri‐methylation of lysine 36 of histone H3, a chromatin modification that is set co‐transcriptionally, provides a quantitative measure of the process of RNA synthesis. We built a linear model that combines H3K36 tri‐methylation with other histone marks and Pol‐II occupancy to predict transcription and to relate it to mRNA levels. This reveals a high correlation between predicted transcription based on chromatin and actual mRNA abundance in both dividing pluripotent cells and terminally differentiated neurons, suggesting that transcription and mRNA levels are tightly linked at different cellular stages. These findings are consistent with two recent studies comparing direct measures of transcription with mRNA abundance (Rabani et al, 2011; Schwanhäusser et al, 2011). Furthermore, we investigated the predictive power of histone marks towards ...
BioAssay record AID 755109 submitted by ChEMBL: Antiviral activity against HCV JFH-1 infected in human Huh7.5.1 cells assessed as viral core protein expression at 0.8 to 8 uM after 72 hrs by Western blot analysis.
The presence of angiotensinogen messenger RNA (mRNA) was assessed in total RNA extracted from hepatoma, glioma, neuroblastoma, and glioma-neuroblastoma hybrid cell lines. Total RNA from 1 X 10(7) cells was extracted, transferred to a membrane, and hybridized with a 32P-labeled, full-length (1650-base pair) rat angiotensinogen complementary DNA (cDNA). Angiotensinogen RNA sequences could be definitively detected only in hepatoma cells. Steroids were used in an attempt to increase the angiotensinogen mRNA level. Dexamethasone (2 X 10(-6) M) or 17 beta-estradiol (1 X 10(-7) M) was added to the cultures 18 to 24 hours prior to harvest. Dexamethasone treatment of the hepatoma cells resulted in a large increase in angiotensinogen mRNA, whereas estradiol had no effect. Steroids failed to induce detectable levels of angiotensinogen mRNA in total RNA from the other cell lines. That the RNA was intact was ensured by hybridizing duplicate Northern blots to a 32P-labeled actin cDNA. Actin mRNA sequences ...
The extent of intratumoral heterogeneity, the subclonal structures and the mechanisms of treatment-induced clonal selection by cisplatin was investigated in the squamous cell carcinoma cell line model FaDu. We picked 96 single cell-derived clones from the cisplatin-sensitive parental FaDu cell line. After expansion as separate cultures, these clones were tested for their sensitivity to CDDP. By this approach, we isolated individual cell clones that were primarily resistant (clones 5 & 78) and others that showed high sensitivity to CDDP (clones 46 & 54). Basal mRNA expression levels associated with CDDP sensitivity / resistance were determined in two independent microarray analyses.
mRNA stability is modulated by elements in the mRNA transcript and their cognate RNA binding proteins. Poly(U) binding protein 1 (Pub1) is a cytoplasmic Saccharomyces cerevisiae mRNA binding protein that stabilizes transcripts containing AU-rich elements (AREs) or stabilizer elements (STEs). In a yeast two-hybrid screen, we identified nuclear poly(A) binding protein 2 (Nab2) as being a Pub1-interacting protein. Nab2 is an essential nucleocytoplasmic shuttling mRNA binding protein that regulates poly(A) tail length and mRNA export. The interaction between Pub1 and Nab2 was confirmed by copurification and in vitro binding assays. The interaction is mediated by the Nab2 zinc finger domain. Analysis of the functional link between these proteins reveals that Nab2, like Pub1, can modulate the stability of specific mRNA transcripts. The half-life of the RPS16B transcript, an ARE-like sequence-containing Pub1 target, is decreased in both nab2-1 and nab2-67 mutants. In contrast, GCN4, an STE-containing ...
Mature messenger RNA, often abbreviated as mature mRNA is a eukaryotic RNA transcript that has been spliced and processed and is ready for translation in the course of protein synthesis. Unlike the eukaryotic RNA immediately after transcription known as precursor messenger RNA, mature mRNA consists exclusively of exons and has all introns removed. Mature mRNA is also called "mature transcript", "mature RNA" or "mRNA". The production of a mature mRNA molecule occurs in 3 steps: During capping, a 7-methylguanosine residue is attached to the 5-terminal end of the primary transcripts. This is otherwise known as the GTP or 5 Cap, and is used for the stability and attachment point for ribosomes. In polyadenylation, a poly-adenosine tail of about 200 adenylate residues is added by a nuclear polymerase post-transcriptionally. This is known as a Poly-A tail and is used for stability and guidance, so that the mRNA can exit the nucleus and find the ribosome. RNA splicing removes the non-coding RNA ...
The embryonic and postnatal expression of 13 GABAA receptor subunit genes in the rat CNS was studied by in situ hybridization. Each transcript exhibited a unique regional and temporal developmental expression profile. For example, in both embryonic and early postnatal cortex and thalamus, expression of the alpha 2, alpha 3, alpha 5, and beta 3 mRNAs was pronounced. In particular, the alpha 5 gene expression underwent a prominent peak in early brain. Subsequently, the thalamocortical expression of these four genes substantially diminished and was superseded in the adult by the alpha 1, alpha 4, beta 2, and delta subunit mRNAs. Similarly, gamma 1 and gamma 3 gene expression also dropped markedly during development, their initial stronger expression being restricted to relatively few structures. In contrast, gamma 2 gene expression was widespread and mostly remained constant with increasing age. The medial septum and globus pallidus were regions expressing few subunits in both early postnatal and ...
AU-rich elements (AREs) in 3-untranslated regions of mRNAs confer instability. They target mRNAs for rapid deadenylation and degradation and may enhance decapping. The p38 MAPK pathway stabilizes many otherwise unstable ARE-containing mRNAs encoding proteins involved in inflammation; however, the mRNA decay step(s) regulated by the signaling pathway are unknown. To investigate whether it regulates deadenylation or the decay of the mRNA body, we used a tetracycline-regulated beta-globin mRNA reporter system to transcribe pulses of mRNA of uniform length. We measured on Northern gels the migration of reporter mRNAs isolated from cells transfected only with reporter plasmid or co-transfected with an active mutant of MAPK kinase-6, and treated either with or without the p38 MAPK inhibitor SB 203580. Differences in migration were shown by RNase H mapping with oligo(dT) to be due to poly(A) shortening. Insertion of an ARE into the beta-globin reporter mRNA promoted rapid deadenylation and decay of hypo
Some work on this problem has been done. A fairly recent reference (with abstract) is given below. Authors Thanaraj TA. Argos P. Title Protein secondary structural types are differentially coded on messenger RNA. Source Protein Science. Vol 5(10) (pp 1973-1983), 1996. Abstract Tricodon regions on messenger RNAs corresponding to a set of proteins from Escherichia coli were scrutinized for their translation speed. The fractional frequency values of the individual codons as they occur in mRNAs of highly expressed genes from Escherichia coli were taken as an indicative measure of the translation speed. The tricodons were classified by the sum of the frequency values of the constituent codons. Examination of the conformation of the encoded amino acid residues in the corresponding protein tertiary structures revealed a correlation between codon usage in mRNA and topological features of the encoded proteins. Alpha helices on proteins tend to be preferentially coded by translationally fast mRNA regions ...
We determined the effects of supplemental L-carnitine on the insulin-like growth factor (IGF) system in porcine embryonic myoblasts (PEM) from gilts. Forty gilts (BW = 303.6 lb) were allotted to 1 of 4 treatments that were arranged in a 2 × 2 factorial, with main effects of L-carnitine (0 or 50 ppm) and day of gestation (55 or 70). All gilts were fed 3.86 lb/day and a top-dress containing either 0 or 50 ppm of L-carnitine, starting on the first day of breeding and continuing through the allotted gestation length. At d 55 or 70 of gestation, fetuses were removed for isolation of PEM from the hind-limb muscles. Real-time quantitative PCR was used to determine growth factor messenger RNA (mRNA) expression in cultured PEM at 72-, 96-, 120-, and 144-h after plating. Flow cytometry was used to analyze percentage of myogenic cells with a myoblast/myotube specific monoclonal antibody 5.1H11, and for determination of cell cycle stage. There was no treatment differences (P,0.10) for the expression of ...
A regulatory locus in a higher organism has been shown to control a specific messenger RNA activity. The Gur locus in mice regulates the production of kidney beta-glucuronidase messenger RNA activity after induction of the beta-glucuronidase structural gene, Gus, by testosterone. beta-Glucuronidase messenger RNA was assayed by its ability to direct the synthesis of catalytically active murine beta-glucuronidase in Xenopus oocytes.
Functions of the PolyA Tail 1.Promotes mRNA stability - Deadenylation (shortening of the polyA tail) can trigger rapid degradation of the mRNA 2.Enhances translation - promotes recruitment by ribosomes - bound by a polyA-binding protein in the cytoplasm called PAB1 - synergistic stimulation with Cap!
Messenger RNA (mRNA) degradation is an important element of gene expression that can be modulated by alterations in translation, such as reductions in initiation or elongation rates. Reducing translation initiation strongly affects mRNA degradation by driving mRNA toward the assembly of a decapping complex, leading to decapping. While mRNA stability decreases as a consequence of translational inhibition, in apparent contradiction several external stresses both inhibit translation initiation and stabilize mRNA. A key difference in these processes is that stresses induce multiple responses, one of which stabilizes mRNAs at the initial and rate-limiting step of general mRNA decay. Because this increase in mRNA stability is directly induced by stress, it is independent of the translational effects of stress, which provide the cell with an opportunity to assess its response to changing environmental conditions. After assessment, the cell can store mRNAs, reinitiate their translation or, ...
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microRNA: microRNA is a short RNA molecule found in eukaryotic cells. A microRNA molecule has very few nucleotides (an average of 22) compared with other RNAs. microRNAs are well conserved in eukaryotic organisms and are thought to be a vital and evolutionarily ancient component of genetic regulation.. microRNAs are post-transcriptional regulators that bind to complementary sequences on target messenger RNA transcripts (mRNAs), usually resulting in translational repression or target degradation and gene silencing. The human genome may encode over 1000 microRNAs, which may target about 60% of mammalian genes and are abundant in many human cell types.. microRNAs show very different characteristics between plants and metazoans. In plants the microRNA complementarity to its mRNA target is nearly perfect, with no or few mismatched bases. In metazoans, on the other hand, microRNA complementarity typically encompasses the 5 bases 2-7 of the microRNA, the microRNA seed region, and thus one microRNA can ...
Post-transcriptional mechanisms of gene regulation play a prominent role during early development. Because the oocyte and developing embryo go through a phase in which no transcription takes place, gene expression relies on a pool of maternal mRNAs accumulated during oogenesis and is regulated at the level of translation or mRNA stability. It has been shown in several biological systems that poly(A) tail shortening contributes to translational silencing, whereas translational activation requires poly(A) tail extension (Richter, 2000; Tadros and Lipshitz, 2005). Poly(A) tail shortening, or deadenylation, is also the first step in mRNA decay. Subsequent steps occur only after the poly(A) tail has been shortened beyond a critical limit (Meyer et al., 2004; Parker and Song, 2004). Rapid deadenylation of unstable RNAs is caused by destabilizing elements, for example AU-rich elements (AREs) found in the 3′ UTRs of several mRNAs. A number of proteins have been identified that bind to destabilizing ...
Dehydroepiandrosterone (DHEA) is a peroxisome proliferating agent when administered in pharmacological dosages, but it has not been shown to function through the peroxisome proliferator-activated receptor in cell-based assays. Because members of the thyroid hormone/vitamins A and D nuclear receptor subfamily, including PPAR, are known to modulate each others function in gene expression by heterodimerization, we sought to establish whether DHEA and thyroid hormone interact to regulate several of the hepatic and renal enzymes associated with peroxisome proliferation, i.e., peroxisomal beta-oxidation and microsomal NADPH:cytochrome P450 oxidoreductase and the cytochromes P450 4A. In rats administered exogenous T3 to attain a hyperthyroid state, induction of the three isozymes of CYP4A (4A1, 4A2, and 4A3) by DHEA was suppressed , 60-80% at the mRNA level, with induction of CYP4A2 mRNA being completely inhibited. Nuclear run-on transcription assays indicated that this inhibitory effect was regulated ...
TY - JOUR. T1 - Regulation of gene expression during adipocyte differentiation. T2 - a review.. AU - Gaskins, H. R.. AU - Hausman, G. J.. AU - Martin, R. J.. PY - 1989/9. Y1 - 1989/9. N2 - The differentiation of adipose precursor cells is accompanied by the acquisition of adipocyte-specific messenger (m) RNAs allowing characteristic changes in protein composition. The development of methods for cloning and characterizing individual genes has provided the opportunity to study selective gene expression by adipocytes at the molecular level. In this review, the information obtained to date regarding transcriptional and post-transcriptional regulatory mechanisms utilized by adipocytes is summarized. Included are descriptions of conserved DNA sequences found in noncoding regions of adipose genes and of how protein-DNA interactions at these regions are thought to regulate the initiation of transcription. Among the transcription factors implemented in regulation of adipocyte-specific gene expression are ...
(a) Collagen type I mRNA expression was increased in 1 h. A persistent high level was formed in postoperative 24 h to 3 d. Despite being slightly descende
Free Online Library: Integrating miRNA and mRNA Expression Profiling Uncovers miRNAs Underlying Fat Deposition in Sheep.(Research Article, Report) by BioMed Research International; Biotechnology industry High technology industry Adipose tissue Analysis Genetic aspects Adipose tissues Genes Messenger RNA Physiological aspects MicroRNA
HER-2 and EGFR have been shown to act synergistically to transform NIH3T3 cells (18), with HER-2 potentiating the signaling of EGFR through increased affinity of ligand binding to EGFR, suppression of EGFR degradation, and enhancement of EGFR recycling (19-21). These observations suggest that both HER-2 and EGFR may play a role in promoting tumor cell growth in selected breast cancers; therefore, both may represent targets for anti-receptor-targeted therapy. From this perspective, inhibition of both receptors could be more effective cancer therapy than inhibition of either receptor alone. To assess the potential clinical utility of HER-2 and EGFR as molecular markers of responsiveness to lapatinib, we retrospectively evaluated the association between these receptors and clinical outcome in two trials of lapatinib treatment for women with metastatic breast cancer. These trials were designed to treat patients with, respectively, HER-2-positive breast cancer (EGF100151) and HER-2-negative/untested ...
MicroRNA (miRNA) directed gene repression is an important mechanism of posttranscriptional regulation. Comprehensive analyses of how microRNA influence biological processes requires paired miRNA-mRNA expression datasets. However, a review of both GEO and ArrayExpress repositories revealed few such datasets, which was in stark contrast to the large number of messenger RNA (mRNA) only datasets. It is of interest that numerous primary miRNAs (precursors of microRNA) are known to be co-expressed with coding genes (host genes). We developed a miRNA-mRNA interaction analyses pipeline. The proposed solution is based on two miRNA expression prediction methods - a scaling function and a linear model. Additionally, miRNA-mRNA anti-correlation analyses are used to determine the most probable miRNA gene targets (i.e. the differentially expressed genes under the influence of up- or down-regulated microRNA). Both the consistency and accuracy of the prediction method is ensured by the application of stringent
Due to the poor correlation between steady state mRNA levels and protein products, traditional microarray analysis may miss many genes which are regulated primarily at the level of mRNA stability and translation. Posttranscriptional gene regulation is becoming increasingly recognized as an important form of cellular control. The importance of microRNAs and RNA binding proteins (RBPs) is now beginning to be better appreciated. We study the elav (embryonic lethal abnormal vision) family of RBPs, which are paraneoplastic antigens, over-expressed in a variety of malignancies, including breast cancer. Antibodies against elav family members are believed to be cancer-protective. The elav family of RBPs binds to the AU-rich elements (AREs) found in the 3\#8217; untranslated regions (UTRs) of many early-response genes, including proto-oncogenes and cell cycle regulators. HuR, the ubiquitously expressed family member, has been described to play a role in cancer progression. HuR stabilizes and ...
A primary transcript is the single-stranded ribonucleic acid (RNA) product synthesized by transcription of DNA, and processed to yield various mature RNA products such as mRNAs, tRNAs, and rRNAs. The primary transcripts designated to be mRNAs are modified in preparation for translation. For example, a precursor messenger RNA (pre-mRNA) is a type of primary transcript that becomes a messenger RNA (mRNA) after processing. There are several steps contributing to the production of primary transcripts. All these steps involve a series of interactions to initiate and complete the transcription of DNA in the nucleus of eukaryotes. Certain factors play key roles in the activation and inhibition of transcription, where they regulate primary transcript production. Transcription produces primary transcripts that are further modified by several processes. These processes include the 5 cap, 3-polyadenylation, and alternative splicing. In particular, alternative splicing directly contributes to the ...
Untranslated regions (UTRs) are sections of the RNA before the start codon and after the stop codon that are not translated, termed the five prime untranslated region (5 UTR) and three prime untranslated region (3 UTR), respectively. These regions are transcribed as part of the same transcript as the coding region. Several roles in gene expression have been attributed to the untranslated regions, including mRNA stability, mRNA localization, and translational efficiency. The ability of a UTR to perform these functions depends on the sequence of the UTR and can differ between mRNAs. The stability of mRNAs may be controlled by the 5 UTR and/or 3 UTR due to varying affinity for RNA degrading enzymes called ribonucleases and for ancillary proteins that can promote or inhibit RNA degradation. Translational efficiency, including sometimes the complete inhibition of translation, can be controlled by UTRs. Proteins that bind to either the 3 or 5 UTR may affect translation by influencing the ...
The integrated multi-omics analysis provides insights into variation at different gene expression levels during the adaption of modern maize from tropical to temperate regions. Population-specific proteome variation mirrors genetic variation better than mRNA levels, and a class of cis-QTLs were identified that regulate protein abundance with little or no effect on mRNA levels. Thus, the discordance between protein and mRNA levels indicates far greater evolutionary stability of proteome during modern maize breeding. ...
TY - JOUR. T1 - A novel CSF‐1 binding factor in a patient in complete remission following cytotoxic therapy for lymphoma. AU - Baker, A. H.. AU - Cachia, P. G.. AU - Tennant, G. B.. AU - Whittaker, J. A.. AU - White, D.. AU - Stanley, E. R.. AU - Burnett, A. K.. AU - Padua, R. A.. PY - 1995/1. Y1 - 1995/1. N2 - Summary. A novel colony stimulating factor‐1 (CSF‐1) binding factor present in the serum from a patient in remission from lymphoma is described. Radioimmunoassay (RIA) repeatedly failed to detect circulating levels of CSF‐1 in the peripheral blood system of this patient. Molecular analysis showed a normal CSF‐1 gene structure by Southern blot analysis and a 46, XX karyotype by cytogenetic analysis. CSF‐1 mRNA expression in peripheral blood leucocytes was confirmed using reverse transcriptase polymerase chain reaction analysis. Morphological analysis of bone marrow cells was normal and peripheral blood progenitor cell colony assays showed a pattern of growth within the normal ...
Extensive genomic characterization of human cancers presents the problem of inference from genomic abnormalities to cancer phenotypes. To address this problem, we analysed proteomes of colon and rectal tumours characterized previously by The Cancer Genome Atlas (TCGA) and perform integrated proteogenomic analyses. Somatic variants displayed reduced protein abundance compared to germline variants. Messenger RNA transcript abundance did not reliably predict protein abundance differences between tumours. Proteomics identified five proteomic subtypes in the TCGA cohort, two of which overlapped with the TCGA microsatellite instability/CpG island methylation phenotype transcriptomic subtype, but had distinct mutation, methylation and protein expression patterns associated with different clinical outcomes. Although copy number alterations showed strong cis- and trans-effects on mRNA abundance, relatively few of these extend to the protein level. Thus, proteomics data enabled prioritization of ...
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The following description of the experiment is taken from that vignette.. Experimental Data "The investigators in this experiment were interested in the effect of estrogen on the genes in ER+ breast cancer cells over time. After serum starvation of all eight samples, they exposed four samples to estrogen, and then measured mRNA transcript abundance after 10 hours for two samples and 48 hours for the other two. They left the remaining four samples untreated, and measured mRNA transcript abundance at 10 hours for two samples, and 48 hours for the other two. Since there are two factors in this experiment (estrogen and time), each at two levels (present or absent,10 hours or 48 hours), this experiment is said to have a 2x2 factorial design." The table below describes the experimental conditions for each of the eight arrays.. ...
As reported elsewhere,[1] the study of RNA synthesis in vitro in immature duck erythrocytes has revealed the occurrence in the nucleus of fast turning-over RNA of high molecular weight (from 2 X 10^6 to 10^7, or possibly more) and with base composition different from that of ribosomal RNA, and characterized by a high U and relatively low GC content. No relationship of precursor-to-product type -- or, at any rate, not a simple one-appears to exist between this RNA and the messenger RNA fraction associated with cytoplasmic polysomes. In this paper, evidence is presented indicating that this class of nuclear RNA molecules is not exclusive of immature erythrocytes, or in general of nondividing cells undergoing differentiation, but occurs also in exponentially growing cells. ...
subject: discussion on Quantitative Competitive RT-PCR dear netters, I am busy writing a discussion for a report I hope graduate on. So at the moment I am looking for some additional comments and insights on Quantitative Competitive RT-PCR that will help me getting a more thorough discussion. Here are some of the questions I am taking in account: What are the strong and weak points in the QC RT-PCR system (internal control: insertion of approx. 4bp; 5 fluorescent labeled primer; genescan analysis/ABI 370)? What is the best way to validate the method you are trying to set up. And are the results obtained with this method reliable/reproducible enough to get accepted by reviewers. Is there some strong literature discussing the system including the disadvantages of it. Thanks in advance, David E. Sluijs Pathology, Academic Hospital Utrecht. please mail to: ,paspoort at dds.nl ...
Protein synthesis in neuronal dendrites underlies long-term memory formation in the brain. Local translation of reporter mRNAs has demonstrated translation in dendrites at focal points called translational hotspots. Various reports have shown that hundreds to thousands of mRNAs are localized to dendrites, yet the dynamics of translation of multiple dendritic mRNAs has remained elusive. Here, we show that the protein translational activities of two dendritically localized mRNAs are spatiotemporally complex but constrained by the translational hotspots in which they are colocalized. Cotransfection of glutamate receptor 2 (GluR2) and GluR4 mRNAs (engineered to encode different fluorescent proteins) into rat hippocampal neurons demonstrates a heterogeneous distribution of translational hotspots for the two mRNAs along dendrites. Stimulation with s-3,5-dihydroxy-phenylglycine modifies the translational dynamics of both of these RNAs in a complex saturable manner. These results suggest that the ...
http://updepla1srv1.epfl.ch/getprime/ A gene- or transcript-specific primer database for quantitative real-time PCR] This user-friendly plateform uniquely combines and automates several features critical for optimal qPCR primer design. These include the consideration of all gene splice variants to enable either gene-specific (covering the majority of splice variants) or transcript-specific (covering one splice variant) expression profiling, primer specificity validation, automated best primer pair selection according to strict criteria and graphical visualization of the latter primer pairs within their genomic context. GETPrime primers have been extensively validated experimentally demonstrating their high quality and demonstrating high transcript specificity in complex samples. Until now, you could retrieve primers in a high-throughput fashion for all Homo sapiens, Mus musculus, Caenorhabditis elegans, Drosophila melanogaster and Danio rerio genes in ...
http://updepla1srv1.epfl.ch/getprime/ A gene- or transcript-specific primer database for quantitative real-time PCR] This user-friendly plateform uniquely combines and automates several features critical for optimal qPCR primer design. These include the consideration of all gene splice variants to enable either gene-specific (covering the majority of splice variants) or transcript-specific (covering one splice variant) expression profiling, primer specificity validation, automated best primer pair selection according to strict criteria and graphical visualization of the latter primer pairs within their genomic context. GETPrime primers have been extensively validated experimentally demonstrating their high quality and demonstrating high transcript specificity in complex samples. Until now, you could retrieve primers in a high-throughput fashion for all Homo sapiens, Mus musculus, Caenorhabditis elegans, Drosophila melanogaster and Danio rerio genes in ...
BioAssay record AID 350380 submitted by ChEMBL: Induction of apoptosis in human Jurkat E6-1 cells assessed as reduction of Bcl-2 mRNA expression at 85 nM after 6 to 18 hrs by RT-PCR analysis.
Using tissue samples obtained from a previous study, the effect of naproxen on the gene expression profiles of antral mucosal tissue will be assessed. We hypothesize that there will be distinct changes in the gene expression profiles of samples taken from individuals treated with naproxen versus samples taken from individuals treated with placebo ...
Project Title: Combinatorial Control of Gene Expression by MiRNAs and RBPs. Gene expression is elaborately controlled at the post-transcriptional level. Molecularly, this control is accomplished through interactions of small RNAs (microRNAs) and RNA-binding proteins (RBPs) with specific sites on messenger RNAs; these associations affect the stability and translation rates of the mRNAs, altering the protein output. As the cell or its environment changes (for example, in cases of differentiation or stress), the interactions between these factors and mRNAs dynamically rearrange to adjust the expression program. Furthermore, binding of microRNAs or RBPs at nearby sites on the same mRNA allows for additive or synergistic interactions between the regulators themselves, imparting a higher order of combinatorial control. In this way, hundreds of microRNAs and RBPs functioning in mammalian cells set up a post-transcriptional regulatory network.. Our research focuses on understanding the overall mapping ...
Figure 2. The phenotype of FoxC1 depleted gastulae. A:Xenopus FoxC1 pseudoalleles aligned against the reverse complement of the morpholino oligo used in the study using Clustal alignment in Macvector. B: FoxC1 MO (MO) blocks translation of FoxC1 mRNA (FoxC1) in a dose-responsive fashion, but does not block translation of a protected FoxC1 RNA, (MOR-), in an in vitro translation assay. C: HA-tagged Fox1 mRNA was injected alone or together with different doses of FoxC1 MO at the 2-cell stage to confirm a reduction of FoxC1 translation. D-G: Wild type sibling embryos and embryos injected with 40 ng MO (MO), or 40 ng MO and 50 pg MO-R FoxC1 mRNA (MO+RNA) were cultured until early (stage 10), mid (stage 11), and late gastrula (stage 11.5) stages before freezing and analyzing by real time RT-PCR for the expression of Mespo (D), Endodermin (E), Bix4 (F), and CK1ϵ (G). Expression levels are normalized to ODC. H,I: Whole mount in situ hybridization on FoxC1-depleted mid-gastrulae (bottom) as compared to ...
Cancer cells are unique in that they can persist (grow and survive) under stressful conditions, such as hypoxia, chemotherapy, and radiation. Cancer cells do this by re-programing transcriptional and post-transcriptional processes that contribute to cell growth, invasion and survival. These post-transcriptional processes include microRNA-mediated mRNA silencing, mRNA decay, mRNA surveillance and translational repression. The induction of these processes results in the formation of Processing Bodies (P-bodies), dynamic cytoplasmic granules that contain mRNAs, microRNAs, and several mRNA processing enzymes, in the cell. In our lab, we have found that P-bodies are associated with a biological process known as epithelial-to-mesenchymal transition (EMT) in breast cancer cells. EMT promotes the polarization and motility of epithelial cells to undergo biochemical and epigenetic changes to assume a mesenchymal phenotype and often times stem-like properties.. Recent studies indicate a role for tyrosine ...
Differential mRNA expression of prostaglandin receptor subtypes in macrophage activation. Hubbard, N.E.; Lee, Seung-Hyoresearcher; Lim, D.; Erickson, K.L., PROSTAGLANDINS LEUKOTRIENES AND ESSENTIAL FATTY ACIDS, v.65, no.5-6, pp.287 - 294, 2001- ...
Summary. Work in my lab is focusing on understanding how microorganisms manage to rapidly adapt (and even thrive) to sudden changes in their environment. Many pathogenic species have developed very sophisticated mechanisms to efficiently scavenge essential nutrients from the host environment and even evade the immune system.. We hypothesize that this successful rapid adaptation program is underpinned by the ability of the microorganism to very rapidly remodel its gene expression profile. Obviously, transcription factors largely dictate which genes are switched on and off during adaptive responses.. However, it is becoming increasingly clear that post-transcriptional regulation plays a key role in this process by shaping gene expression profiles. Small non-coding RNAs (sRNAs) and RNA-binding proteins (RBPs) are believed to play a crucial role in post-transcriptional regulation by modulating the translation efficiency and stability of mRNA targets. However, for the vast majority their function is ...
Glutathione S-transferase Mu 1 (GSTM1) has been regarded as one of the key enzymes involved in phase II reactions in the liver, because of its high expression level. In this study, we generated mice with disrupted glutathione S-transferase Mu 1 gene (Gstm1-null mice) by gene targeting, and characterized the phenotypes by cytosolic and in vivo studies. The resulting Gstm1-null mice appeared to be normal and were fertile. Expression analyses for the Gstm1-null mice revealed a deletion of Gstm1 mRNA and a small decrease in glutathione S-transferase alpha 3 mRNA. In the enzymatic study, GST activities toward 1,2-dichloro-4-nitrobenzene (DCNB) and 1-chloro-2,4-dinitrobenzene (CDNB) in the liver and kidney cytosols were markedly lower in Gstm1-null mice than in the wild-type control. Gstm1-null mice had GST activities of only 6.1 to 21.0% of the wild-type control to DCNB and 26.0 to 78.6% of the wild-type control to CDNB. After a single oral administration of DCNB to Gstm1-null mice, the plasma ...
How different types of cells in intact tissues regulate their mRNAs in physiologic and disease conditions remains largely unexplored. My research goal is to study mRNA regulation in both physiologic and disease scenarios with a special emphasis on 3-prime UTR-mediated regulation. Cells use 3-prime untranslated regions (3-prime UTRs) as dynamic platforms to regulate mRNA stability, localization and translational efficiency in response to outside stimuli by interacting with regulators such as microRNAs (miRNAs) and RNA-binding proteins (RBPs). Moreover, erroneous 3-prime UTR-mediated mRNA regulation is frequently involved in human disease. One important mechanism to generate mRNA isoforms with distinct 3-prime UTR sequences is through alternative polyadenylation (APA), which has been recently shown to play important roles in fundamental cellular processes including proliferation and differentiation, and also in human disease such as cancer, vascular thrombosis, and neurological disorders. During ...
The message transported through mRNA after a certain amount of time will be degraded and be deleted. This process is called degradation. The cell can easily and quickly changed the protein production in case of any changing needs due to the lifetime of the mRNA. The lifetime of different types of mRNA can be different.The life span of mRNA molecules in the cytoplasm is an important key in determining the pattern of protein synthesis within a cell. Prokaryotic mRNA molecules often are degraded by enzymes within a few minutes of their synthesis and this is one reason as to why prokaryotes can vary their patterns of protein synthesis so quickly in response to changes in their environment. Eukaryotic mRNA, on the other hand, typically survives for hours, days, or for some instances, weeks. One example of multicellular mRNA is hemoglobin polypeptides which, in the process of developing red blood cells which are unusually stable, these long-lived mRNAs are translated repeatedly in the cell. Research ...
A translation lookaside buffer (TLB) stores translation entries. The translation entries include a virtual address, a physical address and a memory local/not-local flag. When a processor is in a low power/local memory mode a virtual address is received. A matching translation entry has a local/not-local flag. Upon the local/not-local flag indicating the physical address of the matching translation entry being outside the local memory, an out-of-access-range memory access exception is generated.
Rocklands Immunohistochemistry Studies (IHC) provide confidential high-quality target localization data through wide services. Option to outsource IHC.
(KudoZ) English to Portuguese translation of marked down-regulation: regulação descendente marcante, regulação descendente acentuada [Medical (general) (Medical)].
Due to its high nuclease resistance and improved stability in biological fluids, circular DNA (cDNA) has been used to fabricate an intracellular mRNA sensing platform. cDNA has turned out to be a good option for highly efficient biosensing and therapeutics in living biological systems.
Selective mRNA turnover is an important mechanism of eukaryotic gene regulation. The expression of proto‐oncogenes, lymphokines and cytokines is usually transient, requiring rapid mRNA removal through destabilization following the cessation of transcription. AU‐rich elements (AREs) located in their 3′ untranslated regions (3′ UTRs) comprise a major class of cis‐elements that target these mRNAs for rapid degradation (Caput et al., 1986; Shaw and Kamen, 1986; for reviews, see Belasco and Brawerman, 1993; Chen and Shyu, 1995). Loss of this negative regulatory control conferred by AREs has been shown to be associated with transforming phenotypes (Miller et al., 1984; Meijlink et al., 1985; Lee,W. et al., 1988). Besides mRNAs, small nuclear RNAs (snRNAs) can also be targeted by AREs for rapid degradation, presumably through similar decay pathway(s) (Fan et al., 1997). ARE‐mediated decay may also be differentially regulated. For instance, in a monocyte tumor cell line, c‐fos mRNA is ...
Protein synthesis is a vital cellular process that regulates growth and metabolism. It is controlled via signaling networks in response to environmental changes, including the presence of nutrients, mitogens, or starvation. The phosphorylation state of proteins involved in translation initiation is a limiting factor that regulates the formation or activity of translational complexes. In cancer cells, hyperactivated signaling pathways influence translation, allowing uncontrolled growth and survival. In addition, several components of translation initiation have been found to be mutated, posttranslationally modified, or differentially expressed, and some act as oncogenes in cancer cells. Translational alterations can increase the overall rate of protein synthesis as well as activate regulatory mechanisms leading to the translation of specific messenger RNAs for proteins that promote cancer progression and survival. Many recent studies investigating such mechanisms have produced ideas for ...