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)

A genomic clone, which includes the CTP: phosphocholine cytidylyltransferase (CCT) open reading frame and its 5'- and 3'- flanking non-coding regions, has been isolated from Arabidopsis thaliana and sequenced. The CCT gene is approximately 3.0 kb in length and contains 8 exons interrupted by 7 introns, which range from 74 to 626 nucleotides. All nucleotide sequences for the intron 3' splice sites are consistent with the consensus AG sequence of plant pre-mRNA processing, while the major GT consensus sequence for the 5' splice site is conserved in 5 of 7 introns. Introns 5 and 6 have a minor GC consensus sequence instead. In the 5'-flanking region there are two sequences related to a cold-responsive element found in the cold-inducible promoter of the A. thaliana cor15a gene, plus one gibberellin-response element. The results from reverse transcriptase-PCR indicate that the expression of A. thaliana CCT is regulated by temperature. The expression level of CCT increased after a 30-min treatment at 5 degrees C. When the plants were returned to 22 degrees C, the expression of CCT also decreased to the original level.
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PMID:Structure and expression of a CTP: phosphocholine cytidylyltransferase gene from Arabidopsis thaliana. 1126 28

Retron reverse transcriptases are unusual procaryotic enzymes capable of synthesis of low-molecular-weight DNA by reverse transcription. All of the so-far-described DNA species synthesized by retron reverse transcriptases have been identified as multicopy single-stranded DNA. We have shown that Salmonella enterica serovar Enteritidis is also capable of synthesis of the low-molecular-weight DNA by retron reverse transcriptase. Surprisingly, Salmonella serovar Enteritidis-produced low-molecular-weight DNA was shown to be a double-stranded DNA with single-stranded overhangs (sdsDNA). The sdsDNA was 72 nucleotides (nt) long, of which a 38-nt sequence was formed by double-stranded DNA with 19- and 15-nt single-stranded overhangs, respectively. Three open reading frames (ORFs), encoded by the 4,053-bp plasmid, were essential for the production of sdsDNA. These included an ORF with an unknown function, the retron reverse transcriptase, and an ORF encoding the cold shock protein homologue. This plasmid was also able to confer phage resistance onto the host cell by a mechanism which was independent of sdsDNA synthesis.
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PMID:Low-molecular-weight plasmid of Salmonella enterica serovar Enteritidis codes for retron reverse transcriptase and influences phage resistance. 1129 5

The effectiveness of maintaining prokaryotic RNA in Synechococcus and Pseudomonas cells, fixed in 96% ethanol, 4% paraformaldehyde, or suspended in RNAlater, and held in cold storage for 3 months was compared. Fluorometric determination of the RNA extracted from Synechococcus and Pseudomonas cells indicated that the cell storage treatments tested were equally effective at maintaining their total RNA content. There was not any detectable decrease in the quantity of RNA isolated from the preserved samples during storage. Intact mRNA transcripts of the RuBisCO (rbcL) and nir genes were detected by reverse transcriptase-polymerase chain reaction (RT-PCR) from preserved bacterial cells throughout 3 months of storage. In contrast, RT-PCR failed to amplify the mRNA of the rbcL and nitrite reductase genes in unfixed and/or unpreserved bacterial samples, suggesting that bacterial mRNA can be well maintained during a prolonged storage when cells are preserved properly. In addition, RNAlater is a useful reagent for the storage and maintenance of high quality RNA in unfrozen samples.
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PMID:RNA recovery and detection of mRNA by RT-PCR from preserved prokaryotic samples. 1147 Mar 50

Until now, uncoupling protein 1 (UCP1) was considered as unique to brown adipocytes. It supports a highly regulated uncoupling of oxidative phosphorylation that is associated with diet as well as with non-shivering thermogenesis. Here we report that UCP1 is not specific to brown adipocytes and can be expressed in longitudinal smooth muscle layers. In the uterus, this conclusion was drawn from different convergent data. A specific antibody against mouse UCP1 revealed, in mitochondrial fractions, a protein with the same molecular weight as brown fat UCP1. Sensitive and specific reverse transcriptase-polymerase chain reaction detected a mRNA whose sequence was totally homologous to that of brown fat UCP1 mRNA. Antibody against UCP1 as well as a UCP1 antisense probe specifically stained uterine longitudinal smooth muscles. UCP1 was also expressed in longitudinal smooth muscle of digestive and male reproductive tracts but was never expressed in other types of smooth muscle, including those of arterial vessels. In uterine tract, UCP1 content was increased after cold exposure or beta-adrenergic agonist treatment. It was also up-regulated during the postovulatory period after sexual cycle synchronization. Its content transiently increased during gestation and decreased markedly after birth. These regulations strongly argue about a role for UCP1 in thermogenesis as well as in relaxation of longitudinal smooth muscle layers.
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PMID:Brown fat UCP1 is specifically expressed in uterine longitudinal smooth muscle cells. 1157 62

We evaluated the effect of holding temperature and time between mosquito death and processing mosquito pools for virus detection on our ability to detect West Nile (WN) viral RNA from pools of mosquitoes by reverse transcriptase-polymerase chain reaction (RT-PCR). Pools of 24 uninfected Culex pipiens L. mosquitoes were "spiked" with either a single Cx. pipiens that had been inoculated previously with WN virus or with an uninfected mosquito. These pools were held dry at 20, 4, -20, or -70 degrees C for selected time intervals before all mosquito pools were triturated in TRIzol LS reagent and processed for detection of WN viral RNA. While infectious virus virtually disappeared from pools maintained at 20 degrees C by 48 h after mosquito death, neither holding temperature (20 to -70 degrees C) nor holding period (up to 2 wk) affected detection of WN viral RNA by real-time RT-PCR. These findings suggest that we need not keep mosquitoes chilled to be able to detect WN viral RNA effectively by RT-PCR. This should enhance the feasibility of field-based WN virus surveillance programs where only detection of WN viral RNA is the objective and maintenance of a cold chain may not be possible.
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PMID:Effect of holding conditions on the detection of West Nile viral RNA by reverse transcriptase-polymerase chain reaction from mosquito (Diptera: Culicidae) pools. 1193 Dec 39

Walleye dermal sarcoma virus (WDSV) is a piscine retrovirus that replicates naturally in fish at temperatures near 4 degrees C. The reverse transcriptase (RT) protein from virus particles isolated from walleye tumours was purified and biochemically characterized. Like the RT of the distantly related murine leukaemia virus, WDSV RT sediments as a monomer in the absence of template. It exhibits a K(m) of 22 microM for TTP in an assay with poly(rA) as a template and oligo(dT) as a primer. The enzyme is rapidly inactivated at temperatures greater than 15 degrees C. The ratio of RT activity at 15 degrees C to that at 4 degrees C is similar for WDSV and recombinant human immunodeficiency virus type 1, suggesting that, at least with this template, the fish enzyme is not specially adapted to function more efficiently in the cold.
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PMID:Walleye dermal sarcoma virus reverse transcriptase is temperature sensitive. 1202 51

Necrotizing enterocolitis (NEC) is a common and devastating gastrointestinal disease of premature infants. Because the proinflammatory cytokines IL-18, IL-12, and interferon (IFN)-gamma have been implicated in other diseases of the small intestine, we hypothesized that these cytokines would play an important role in NEC pathogenesis. NEC was induced in newborn rats via enteral feeding with rat milk substitute and asphyxia and cold stress (RMS). Dam-fed, asphyxia- and cold-stressed littermates were used as controls (DF). After 96 h, the distal ileum was removed from all animals and processed to determine expression and localization of IL-18, IL-12, and IFN-gamma using real-time reverse transcriptase PCR and immunohistology. IL-18 and IL-12 mRNA from the RMS group were increased (p < or = 0.05) compared with DF controls, and there was a correlation between increasing IL-18 and IL-12 mRNA levels and progression of tissue damage (r = 0.629 and 0.588, respectively; p < or = 0.05). Immunohistology revealed IL-18 in the cytoplasm of villi and crypt enterocytes and IL-12-positive monocytes/macrophages were increased with disease progression (r = 0.503, p < or = 0.05). No differences in the number of IFN-gamma-positive cells were observed between groups. These data demonstrate up-regulation of IL-18 and IL-12 in experimental NEC and a correlation between production of these proinflammatory cytokines and progression of tissue damage.
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PMID:Up-regulation of IL-18 and IL-12 in the ileum of neonatal rats with necrotizing enterocolitis. 1203 69

Plants respond to environmental stress with a number of physiological and developmental changes. Water deficit is one of the major factors limiting plant growth and development and crop productivity. One response of plants to water deficit is accumulation of abscisic acid (ABA). An increase of ABA is responsible for the induction of many genes, presumably some of which contribute to drought tolerance. Analysis of gene expression in barley seedling shoots by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR) led to the isolation of several drought-, cold- and ABA-induced partial cDNA fragments. Here we extensively characterize one of these cDNAs, designated DD6. First, a larger cDNA was extended from DD6 by 5'-RACE (rapid amplification of cDNA ends). Subsequently, the corresponding gene was isolated by screening a barley BAC library, and the sequences of the transcribed and flanking regions were determined. The deduced amino acid sequence has similarity to an Arabidopsis hypothetical protein and to a human and mouse DNA-binding protein. The corresponding gene, named Srg6 (stress-responsive gene), was mapped in a barley doubled haploid mapping population to chromosome 7H between markers ABC455 and salfp76, within a region that previously has been linked to osmotic adaptation in barley and other grass genomes.
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PMID:Identification and mapping of a putative stress response regulator gene in barley. 1213 5

A 58-year-old man was admitted with generalized lymphadenopathy. On admission, the patient showed polyclonal hyper-gammopathy in a blood examination, positive results in the direct/indirect Coombs test, and an elevated cold agglutinin titer. Autoimmune thrombocytopenia with a high level of platelet-associated immunoglobulin G complicated the patient's condition. An enzyme immunoassay kit for human immunodeficiency virus (HIV) recombinant proteins p24, gp41, and gp36 showed positive results. Western blot analysis showed the presence of antibodies cross-reacting with HIV p24 gag protein. HIV RNA was not detected by means of a reverse transcriptase-polymerase chain reaction assay, so the patient was not an HIV carrier. Angioimmunoblastic T-cell lymphoma (AILT) was diagnosed on the basis of lymph node biopsy specimens. We speculated that in this case some of the numerous subtypes of polyclonal gamma globulin had coincidentally cross-reacted with HIV p24. Cross-reactive phenomena with HIV in patients with systemic lupus erythematosus have been well investigated, but to our knowledge our patient is the first case of such cross-reactivity involving AILT. Physicians should pay close attention to serologic tests to determine whether the patient truly is a viral carrier.
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PMID:Angioimmunoblastic T-cell lymphoma associated with an antibody to human immunodeficiency virus protein. 1295 12

The effect of different physical and chemical factors on the process of transition of persisting mycoplasmas into active virulent state was studied to find out conditions promoting the development of acute infection in the course of chronic infection. The activity of the gene coding the synthesis of protein P1, the main pathogenicity factor, was evaluated in the reverse transcriptase test--PCR under conditions of heat and cold shocks, oxidation stress, varying osmotic pressure. An increased osmotic pressure, heat and cold shocks were shown to induce the transcription activation of the gene coding the synthesis of P1 in the avirulent strain, and consequent restoration of its adhesive properties indicative of virulence. M. pneumoniae avirulent strain was characterized by greater resistance to oxidation stress and to a rise in osmotic pressure, this property requiring further study. Quite probably, M. pneumoniae DNA-binding proteins earlier detected in persisting and avirulent cultures by taking part in the compactization of DNA contribute to the adaptation of mycoplasmas to different stress influences. There are grounds to suggest that M. pneumoniae possess mechanisms regulating the expression of genes, in particular the gene coding the main pathogenicity factor, under the influence of different environmental factors.
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PMID:[Influence of environmental factors on the expression of Mycoplasma pneumoniae gene responsible for the synthesis of protein P1 adhesion]. 1296 70


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