Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.49 (reverse transcriptase)
31,746 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The role of the stress response regulator sigma(B) (encoded by sigB) in directing the expression of selected putative and confirmed cold response genes was evaluated using Listeria monocytogenes 10403S and an isogenic DeltasigB mutant, which were either cold shocked at 4 degrees C in brain heart infusion (BHI) broth for up to 30 min or grown at 4 degrees C in BHI for 12 days. Transcript levels of the housekeeping genes rpoB and gap, the sigma(B)-dependent genes opuCA and bsh, and the cold stress genes ltrC, oppA, and fri were measured using quantitative reverse transcriptase PCR. Transcriptional start sites for ltrC, oppA, and fri were determined using rapid amplification of cDNA ends PCR. Centrifugation was found to rapidly induce sigma(B)-dependent transcription, which necessitated the use of centrifugation-independent protocols to evaluate the contributions of sigma(B) to transcription during cold shock. Our data confirmed that transcription of the cold stress genes ltrC and fri is at least partially sigma(B) dependent and experimentally identified a sigma(B)-dependent ltrC promoter. In addition, our data indicate that (i) while sigma(B) activity is induced during 30 min of cold shock, this cold shock does not induce the transcription of sigma(B)-dependent or -independent cold shock genes; (ii) sigma(B) is not required for L. monocytogenes growth at 4 degrees C in BHI; and (iii) transcription of the putative cold stress genes opuCA, fri, and oppA is sigma(B) independent during growth at 4 degrees C, while both bsh and ltrC show growth phase and sigma(B)-dependent transcription during growth at 4 degrees C. We conclude that sigma(B)-dependent and sigma(B)-independent mechanisms contribute to the ability of L. monocytogenes to survive and grow at low temperatures.
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PMID:SigmaB-dependent and sigmaB-independent mechanisms contribute to transcription of Listeria monocytogenes cold stress genes during cold shock and cold growth. 1767 28

Whole-genome microarray experiments were performed to define the Listeria monocytogenes cold growth regulon and to identify genes differentially expressed during growth at 4 and 37 degrees C. Microarray analysis using a stringent cutoff (adjusted P < 0.001; >/=2.0-fold change) revealed 105 and 170 genes that showed higher transcript levels in logarithmic- and stationary-phase cells, respectively, at 4 degrees C than in cells grown at 37 degrees C. A total of 74 and 102 genes showed lower transcript levels in logarithmic- and stationary-phase cells, respectively, grown at 4 degrees C. Genes with higher transcript levels at 4 degrees C in both stationary- and log-phase cells included genes encoding a two-component response regulator (lmo0287), a cold shock protein (cspL), and two RNA helicases (lmo0866 and lmo1722), whereas a number of genes encoding virulence factors and heat shock proteins showed lower transcript levels at 4 degrees C. Selected genes that showed higher transcript levels at 4 degrees C during both stationary and log phases were confirmed by quantitative reverse transcriptase PCR. Our data show that (i) a large number of L. monocytogenes genes are differentially expressed at 4 and 37 degrees C, with more genes showing higher transcript levels than lower transcript levels at 4 degrees C, (ii) L. monocytogenes genes with higher transcript levels at 4 degrees C include a number of genes and operons with previously reported or plausible roles in cold adaptation, and (iii) L. monocytogenes genes with lower transcript levels at 4 degrees C include a number of virulence and virulence-associated genes as well as some heat shock genes.
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PMID:Microarray-based characterization of the Listeria monocytogenes cold regulon in log- and stationary-phase cells. 1772 Aug 27

A small heat shock protein gene (hsp22.4) was cloned from Chaetomium globosum using rapid amplification of cDNA ends (RACE). The 986-bp full-length hsp22.4 cDNA contains a 609-bp open reading frame encoding a 202-amino-acid protein with an estimated molecular mass of 22.4 kDa. The hsp22.4 gene was amplified using specific primers in the 5' and 3' untranslated regions of the hsp22.4 cDNA. The temporal expression of hsp22.4 was measured in C. globosum by real-time reverse transcriptase-polymerase chain reaction after exposure to heat, cold, Na(2)CO(3), and NaCl. The expression of hsp22.4 was induced by heat and Na(2)CO(3) treatment and inhibited by cold and NaCl treatment. The hsp22.4 gene was inserted into pYES2 containing the inducible GAL1 promoter and transferred into yeast (Saccharomyces cerevisiae) for expression. The hsp22.4 transgenic yeast displayed significantly greater resistance to heat and Na(2)CO(3) stresses than control (yeast cells transformed with empty pYES2), suggesting that the expression of hsp22.4 gene confers not only heat tolerance but also significant alkali (Na(2)CO(3)) stress tolerance.
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PMID:A heat shock protein gene (hsp22.4) from Chaetomium globosum confers heat and Na2CO3 tolerance to yeast. 1794 Jul 62

Shellstock refrigeration after harvesting is recommended to prevent further increases in Vibrio vulnificus numbers in oysters, but it could potentially induce a cold shock response in this bacterium. V. vulnificus was incubated at 35, 25, 20, and 15 degrees C and then subjected to 7.2 and 4 degrees C for 1 week. A cold-adaptation response that enhanced cell culturability was observed when cells were incubated at 15 degrees C prior to cold shock at 7.2 degrees C. In vitro cold shock gene expression was analyzed by reverse transcriptase PCR (RT-PCR). The expression of cold shock genes csp1 and csp5 (homologous genes to cspA and cspV) remained constant, despite cold shock. However, the transcript of csp3 was constitutively expressed before and after cold shock, with a few exceptions. The synthesis of csp3 mRNA in V. vulnificus C7184Tr (an avirulent strain) was induced only after 15 degrees C incubation and cold shock at 4 degrees C. The expression of csp4 was repressed after cold shock. Our data showed that the csp(s) tested in this study are not cold inducible. The transcripts of two oxidative stress-related genes, oxyR and katG, showed different induction patterns among strains after cold shock, suggesting that V. vulnificus cells encountered oxidative stress during cold shock.
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PMID:Gene expression of cold shock and other stress-related genes in Vibrio vulnificus grown in pure culture under shellstock temperature control conditions. 1823 77

Surveillance for avian influenza A viruses in wild bird populations is often limited by requirements for a cold chain from time of specimen collection, by availability of ultra-low temperature specimen storage within a few hours or days, and by laborious classical virologic procedures. Successful storage of specimens in preservatives at ambient temperature and subsequent detection of RNA by reverse transcriptase-polymerase chain reaction (RT-PCR) would assist in helping influenza surveillance efforts become more widespread in remote areas, as well as more timely and inexpensive. Here, we describe bird feces spiked with influenza A virus preserved with guanidine and commercial buffers or alcohols at ambient temperature and analyzed by RT-PCR protocols. Virus-specific RT-PCR products of, at most, 206 bp were recovered for samples preserved with alcohols and up to 521 bp for samples preserved with guanidine or commercial buffers. These results suggest that this approach is feasible in the field and that preserved specimens might be better assayed molecularly when preserved in guanidine or commercial buffers.
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PMID:Effect of preservative on recoverable RT-PCR amplicon length from influenza A virus in bird feces. 1825 9

The early light-induced proteins (ELIPs) are nuclear-encoded, light stress-induced proteins located in thylakoid membranes and related to light-harvesting Chl a/b-binding proteins. Recent evidence from physiological and genetic (mutant) studies supports a photoprotective function for ELIPs, particularly when green tissues are exposed to high light intensities at suboptimal temperatures. Broad-leaved evergreens belonging to genus Rhododendron are often exposed to a combination of low temperatures and high light in their natural habitat as the understory plants in deciduous forests and, therefore, are expected to employ photoprotective strategies during overwintering phase. Here we report analysis and characterization of previously identified ELIP expressed sequence tags (ESTs) from winter-collected Rhododendron catawbiense leaves. 5' or 3' rapid amplification of complementary DNA ends (RACEs) coupled with bioinformatic analyses were used to identify seven unique ELIPs from the 40 ESTs and were designated as RcELIP1-RcELIP7. Phylogenetic analysis revealed separate clustering of ELIP homologs from lower plants, monocots and eudicots (including RcELIPs) and further indicated an evolutionary divergence of ELIPs among angiosperms and gymnosperms. To gain insights into the cold acclimation (CA) physiology of rhododendrons, relative and absolute quantitative expression of RcELIPs was examined during seasonal CA of R. catawbiense leaves using real time reverse transcriptase-polymerase chain reaction. All seven RcELIPs were distinctly upregulated during the CA. It is postulated that RcELIPs expression constitutes an adaptive response to cold and high light in winter-adapted rhododendron leaves and perhaps plays a key role in the protection of photosynthetic apparatus from these stresses.
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PMID:Phylogenetic analysis and seasonal cold acclimation-associated expression of early light-induced protein genes of Rhododendron catawbiense. 1825 69

The 2003 pandemic of Severe Acute Respiratory Syndrome (SARS) profiled the ability of modern diagnostic microbiology and molecular biology to identify, isolate and characterize, within weeks, a previously unknown viral infectious pathogen. The culprit, SARS coronavirus (SARS-CoV), was detected in patient specimens by traditional cell culture using an unusual cell line for respiratory viruses, Vero E6, and by reverse transcriptase polymerase chain reaction (RT-PCR) targeting the polymerase 1 B region of the genome. In addition, serologic assays were rapidly developed, and the genome of this large virus was sequenced within one month of its spread to North America. At the present time, diagnostics have progressed to the point that RT-PCR has a sensitivity approaching 80% within the first few days of onset of illness, while serology has a sensitivity close to 100% on convalescent sera taken >21 days after illness onset. Viral culture remains a method confined to biosafety level III laboratories. The specificity of RT-PCR and serology remains to be conclusively defined, but in most studies to date seems to be >90%. Serologic cross-reactivity with human coronaviruses causing the common cold may be a problem with some serologic assays. The early development of SARS-CoV diagnostics is now being replaced by refinement and optimization of these assays. Although at the present time we do not have a test that will definitively rule in or rule out SARS at the time of initial presentation of a patient with a respiratory infection, modifications of existing assays will hopefully result in our ability to make this diagnosis with a high degree of accuracy in the future.
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PMID:The laboratory diagnosis of severe acute respiratory syndrome: emerging laboratory tests for an emerging pathogen. 1845 11

Expression profiles of nine rice heat shock protein genes (OsHSPs) were analyzed by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR). The nine genes exhibited distinctive expression in different organs. Expression of nine OsHSP genes was affected differentially by abiotic stresses and abscisic acid (ABA). All nine OsHSP genes were induced strongly by heat shock treatment, whereas none of them were induced by cold. The transcripts of OsHSP80.2, OsHSP71.1 and OsHSP23.7 were increased during salt tress treatment. Expression of OsHSP80.2 and OsHSP24.1 genes were enhanced while treated with 10% PEG. Only OsHSP71.1 was induced by ABA while OsHSP24.1 was suppressed by ABA. These observations imply that the nine OsHSP genes may play different roles in plant development and abiotic stress responses.
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PMID:Expression analysis of nine rice heat shock protein genes under abiotic stresses and ABA treatment. 1913 78

Low temperature is a major environmental stress for plants. Many important cultivated crops have limited capacity to survive below freezing/subfreezing temperatures. Low inorganic phosphate (Pi) is reportedly important in triggering cold acclimatization. SPX (SYG1/Pho81/XPR1: SYG1, suppressor of yeast gpal; Pho81, CDK inhibitor in yeast PHO pathway; XPR1, xenotropic and polytropic retrovirus receptor) domain proteins have been shown to be involved in the phosphate-related signal transduction and regulation pathways. Recently, Arabidopsis AtSPX family genes have been found to possess diverse functions in plant tolerance to phosphorus starvation, and OsSPX1 is involved in phosphate homeostasis in rice and optimizes growth under phosphate-limited conditions through a negative feedback loop. In this study, our phylogenetic and gene expression profiling approaches identified six rice OsSPX genes up-regulated during cold stress. Transgenic tobacco plants with constitutive expression of OsSPX1 were more tolerant to cold stress than were wild-type plants, and showed better seedling survival and reduced cellular electrolyte leakage. In addition, there was decreased total leaf Pi content and accumulation of free proline and sucrose in transgenic tobacco plants during cold stress. To further establish a cause-and-effect relationship between intracellular Pi level and cold acclimatization in transgenic plants, we generated transgenic Arabidopsis plants with constitutive expression of OsSPX1. Cold stress resulted in reduced leaf Pi levels in Arabidopsis transgenic relative to wild-type plants. From real-time reverse transcriptase-polymerase chain reaction analysis, several Pi starvation-related genes, such as AtSPX1 (orthologue of OsSPX1), PHO2, PLDZ2 and ATSIZ1, showed differential expression between wild-type and transgenic plants during cold stress. Our results indicate that OsSPX1 may play an important role in linking cold stress and Pi starvation signal transduction pathways.
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PMID:Increased expression of OsSPX1 enhances cold/subfreezing tolerance in tobacco and Arabidopsis thaliana. 1950 76

The hydroxysteroid dehydrogenase HSD1, identified in the proteome of oil bodies from mature Arabidopsis seeds, is encoded by At5g50600 and At5g50700, two gene copies anchored on a duplicated region of chromosome 5. Using a real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) approach, the accumulation of HSD1 mRNA was shown to be specifically and highly induced in oil-accumulating tissues of maturing seeds. HSD1 mRNA disappeared during germination. The activity of HSD1 promoter and the localization of HSD1 transcripts by in situ hybridization were consistent with this pattern. A complementary set of molecular and genetic analyses showed that HSD1 is a target of LEAFY COTYLEDON2, a transcriptional regulator able to bind the promoter of HSD1. Immunoblot analyses and immunolocalization experiments using anti-AtHSD1 antibodies established that the pattern of HSD1 deposition faithfully reflected mRNA accumulation. At the subcellular level, the study of HSD1:GFP fusion proteins showed the targeting of HSD1 to the surface of oil bodies. Transgenic lines overexpressing HSD1 were then obtained to test the importance of proper transcriptional regulation of HSD1 in seeds. Whereas no impact on oil accumulation could be detected, transgenic seeds exhibited lower cold and light requirements to break dormancy, germinate and mobilize storage lipids. Interestingly, overexpressors of HSD1 over-accumulated HSD1 protein in seeds but not in vegetative organs, suggesting that post-transcriptional regulations exist that prevent HSD1 accumulation in tissues deprived of oil bodies.
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PMID:Regulation of HSD1 in seeds of Arabidopsis thaliana. 1954 45


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