Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:2.7.7.48 (transcriptase)
 9,479 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Porphyromonas gingivalis is a gram-negative bacterium that is associated with periodontitis. It has been hypothesized that destruction of bone and periodontal connective tissue is associated with colonization of the subgingival crevicular space by P. gingivalis, although how these bacteria overcome innate host defenses is largely unknown. To examine the early cellular and molecular events of P. gingivalis interaction with host tissues, we compared lipopolysaccharide (LPS) isolated from this bacterium with Escherichia coli LPS, a potent inflammatory mediator, in a mouse model of acute inflammation. In these studies, mice were given intramuscular injections of either P. gingivalis LPS or E. coli LPS and then sacrificed after 4 h. Reverse transcriptase-PCR analysis showed that expression of mRNAs for E- and P-selectins was higher in E. coli LPS-injected muscles than in P. gingivalis LPS-injected or control phosphate-buffered-saline-injected muscles. Similarly, monocyte chemotactic protein 1 and fibroblast-induced cytokine mRNAs were expressed in E. coli LPS-injected muscles whereas their expression was reduced or absent in P. gingivalis LPS-injected samples. These results were confirmed by in situ hybridization whereby stronger hybridization for selectin mRNAs was observed in the endothelium of capillaries from E. coli LPS-injected samples than in that from P. gingivalis LPS-injected muscles. In addition, many monocytes expressing monocyte chemotactic protein 1 mRNA and polymorphonuclear leukocytes expressing fibroblast-induced cytokine mRNA were observed in E. coli LPS-injected muscles whereas only a few cells were identified in P. gingivalis LPS-injected muscles. These results demonstrate that compared with E. coli, P. gingivalis has a low biologically reactive LPS as measured by its weak activation of inflammation. This may allow P. gingivalis to evade innate host defense mechanisms, resulting in colonization and chronic disease.
Infect Immun 1995 Dec
PMID:Porphyromonas gingivalis lipopolysaccharide is poorly recognized by molecular components of innate host defense in a mouse model of early inflammation. 759 Nov 24

Astroviruses cause outbreaks of diarrhea in children attending day care centers (DCCs). Reverse transcriptase-polymerase chain reaction (RT-PCR) was compared with EIA detection of astrovirus in stool specimens to characterize further the molecular epidemiology of an outbreak of astrovirus-associated gastroenteritis. Three hundred sixty-eight stool specimens collected prospectively from 36 children enrolled in a DCC during an 11-week outbreak of diarrhea were evaluated by EIA and RT-PCR. Astrovirus was detected in 32% of specimens by RT-PCR versus 10% by EIA (P < .001) and in 89% of children by RT-PCR versus 50% by EIA. The median duration of astrovirus excretion episodes detected by EIA was 1.5 days versus 4 days by RT-PCR (P = .06). Astrovirus was excreted for prolonged periods by immunocompetent children during this outbreak. RT-PCR was more sensitive than EIA for detection of astrovirus in stool specimens and redefined the epidemiology of astrovirus infection in this setting.
J Infect Dis 1995 Dec
PMID:Virologic features of an astrovirus diarrhea outbreak in a day care center revealed by reverse transcriptase-polymerase chain reaction. 759

The nucleotide sequence of the 5'-end of feline calicivirus (FCV) Japanese F4 strain genome was determined. This region had 5311 bases and contained a large open reading frame (ORF1) encoding the non-structural proteins. The nucleotide sequence of the ORF1 region was highly conserved as compared with that of FCV F9 strain. When the deduced amino acid sequence of the ORF1 was compared with those of FCV F9 and CFI strains, the sequence was also highly conserved (88.9% and 88.8%, respectively). Functional motifs of the non-structural proteins were common to these strains. There were 2C polypeptide-, 3C cysteine protease- and 3D RNA-dependent RNA polymerase-like regions. The N-terminal region of 2C-like region continued upstream from the region identified by Neill [Virus Res. 17: 145-160]. Furthermore, the presence of 2B-like region was suggested in the upper stream of the 2C-like region, although the function of the region is unknown. When Kyte and Dolittle hydrophobicity profiles of the predicted amino acid sequences of the ORF1s of FCV F4 and F9 were computed and compared, both the profiles had striking similarities. In the region between residues 950-1000, there was a high rate of basic amino acid residues, suggesting that the polypeptide in this region of FCV may have a nucleic acid-binding function.
J Vet Med Sci 1994 Dec
PMID:The molecular cloning and sequence of an open reading frame encoding for non-structural proteins of feline calicivirus F4 strain isolated in Japan. 769 98

To study the regulation of mucin synthesis in canine tracheal epithelial cells, it is desirable to establish a cell line which synthesizes mucin continuously. We adopted the approach of immortalizing canine tracheal epithelial cells using a vector encoding the human papillomavirus (type 18) E6 and E7 genes. The E6 and E7 genes are essential and sufficient for the immortalization of human genital keratinocytes, as well as human tracheal epithelium. Primary epithelial cells from dog trachea were transfected with a vector containing HPV18 genes E6 and E7. The resultant cells (CT1) were cloned and maintained in selective medium supplemented with growth factors and hormones. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis indicated the expression of the canine tracheal mucin (CTM) mRNA in these cells. The half-life of the CTM mRNA was found to be 45-60 min. Incorporation of labelled precursor (glucosamine) indicated that high-molecular-weight mucin glycoprotein was synthesized by these immortalized cells, which reacted with the antiserum to the native CTM. Equilibrium gradient centrifugation analysis showed that the buoyant density of the mucin synthesized in CT1 cells (1.486 g/ml) was similar to the reported value for native CTM (1.5 g/ml). Mucin which was isolated from immortalized cells was not a proteoglycan as chondroitinase treatment had no effect. These results suggest that CT1 cells synthesize a mucin glycoprotein which exhibits properties similar to native CTM. When characterized by immunostaining with a pool of monoclonal antibodies, these cells showed common epithelial antigens related to keratin expression. The CT1 cell line represents a unique resource for studying mucin biosynthesis and regulation.
Glycobiology 1994 Dec
PMID:Mucin synthesis in immortalized canine tracheal epithelial cells. 773 45

15-deoxyspergualin (DSG)-treated BALB/c spleen cells showed increased spontaneous proliferation and increased alloreactive mixed lymphocyte reactions (MLRs) when a 3-h treatment was carried out. However, when spleen cells were treated with DSG for 5 days without washing out DSG, decreased spontaneous proliferation was observed, although alloreactive MLRs against C3H/He and C57BL/6 alloantigens were increased. In contrast, cyclosporin A (CsA) induced markedly decreased alloreactive MLRs. Decreased concanavalin A (Con A)- and pokeweed mitogen (PWM)-induced responses were observed in spleen cells treated with DSG for 3 h; whereas increased phytohemagglutinin (PHA)-induced responses were observed. On the other hand, increased Con A- and PHA-induced responses were observed in spleen cells treated with DSG for 2 days, whereas PWM-induced responses were decreased. CsA-treatment induced markedly decreased mitogen-induced responses. These results suggest that the immunosuppressive mechanism of DSG differs from that of CsA. Reverse transcriptase-polymerase chain reaction (RT-PCR) method showed that interleukin 1 beta (IL-1 beta), IL-2, IL-3, IL-4, IL-5, and leukemia inhibitory factor (LIF) mRNA expression in DSG-treated spleen cells were increased by Con A stimulation, thus indicating that DSG modulates cytokine gene expression and inducing immunosuppressive mechanisms different from CsA.
Immunobiology 1994 Dec
PMID:Effects of 15-deoxyspergualin on proliferative responses and cytokine gene expression in vitro. 775 Sep 88

Endothelial cells (ECs) cultured from human umbilical vein were exposed to medium flow in a flow-loading chamber, and changes in thrombomodulin (TM) expression were examined by flow cytometry and enzyme linked immunosorbent assay with monoclonal antibody. The expression of TM antigen was increased time- and shear stress-dependently by flow, and when exposed to a shear stress of 15 dynes/cm2 for 24 hr, it increased to approximately 200% of the stationary control level. Reverse transcriptase-polymerase chain reaction showed that TM mRNA levels in ECs also increased in response to flow. TM mRNA began to increase one hour after the application of shear stress of 15 dynes/cm2 and reached a maximum (approximately 330% of stationary control) after eight hours. These results, demonstrating an up-regulating effect of flow on TM expression in ECs, suggest that shear stress may be an important modulator of intravascular blood coagulation.
Biochem Biophys Res Commun 1994 Dec 15
PMID:Fluid shear stress increases the expression of thrombomodulin by cultured human endothelial cells. 780 68

Abortive cycling features transcription initiation by RNA polymerase in both prokaryote and eukaryote. It is known that T7 RNA polymerase produces abortive transcripts up to eight ribonucleotides in length depending on the initial sequence of the DNA message. On the other hand, T7 RNA polymerase initiates DNA replication from the T7 primary origin by synthesizing primers. And the shortest primer from the phi l.lB promoter in the primary origin also seems to be eight ribonucleotides in length. Therefore, it is likely that the longest abortive transcript serves as the shortest primer for T7 DNA replication from the primary origin. Considering that promoters often exist in DNA replication origins for example, E. coli oriC and many eukaryotic origins, the early DNA replication system appears to have taken advantage of the abortive cycling of RNA-dependent RNA polymerase that already existed before the emergence of DNA world. The evolutionary primitive RNA polymerase could do both transcription and priming of DNA replication. Accordingly, abortive cycling would play an important role in evolution at the emergence of DNA world. The priming activity of the primitive RNA polymerase would be taken over by primase later, which seems to be a specialized RNA polymerase for abortive cycling.
J Mol Evol 1994 Dec
PMID:Evolutionary role of abortive transcript as a primer for DNA replication. 780 50

Studies of RNA replication among the positive-strand RNA animal viruses have been hindered by the apparent inability of their RNA-dependent RNA polymerases to initiate replication on the corresponding negative-sense RNAs. However, here I report that in the case of the nodavirus flock house virus (FHV), which has a bipartite positive-sense RNA genome, the viral RNA replicase can replicate a negative-sense transcript of the genome segment that encodes the viral capsid proteins. For this work, the FHV replication cycle was experimentally reconstructed in baby hamster kidney cells that were transfected with specialized transcription plasmids designed to direct the synthesis of RNAs which corresponded closely to the two genome segments of FHV. The RNA replicase encoded by the larger genome segment could utilize either the positive or the negative strand of the smaller segment as a template, and it catalyzed RNA replication to produce similar RNA products in the two situations. Surprisingly, studies of the nucleotide sequences that were required for replication showed that the 3' end of the negative-strand RNA contained only a minimal cis-acting signal. The success of these experiments will facilitate further studies of the cis- and trans-acting factors involved in the recognition and replication of negative-sense RNA in this system.
Proc Natl Acad Sci U S A 1994 Dec 20
PMID:Replication of the genomic RNA of a positive-strand RNA animal virus from negative-sense transcripts. 780 56

LIM-homeodomain proteins are important in cell lineage specification and possibly mediate transcriptional processes in eukaryotes. During the screening of a mouse pituitary cDNA library, we isolated a partial cDNA coding for a novel gene product that exhibited a predicted amino-terminal sequence similar to the homeobox of LIM-homeodomain-containing proteins. Reverse transcriptase-polymerase chain reactions (RT-PCR) performed on mouse pituitary mRNA using degenerate oligonucleotides based on the conserved LIM-domain sequences, allowed the extension of the 5' end of the sequence. The composite 2.2-kb cDNA structure predicts a 400-amino-acid-long novel mouse (m) protein, called mLIM-3. This name was chosen since within the 59-amino-acid homeodomain, it exhibits 97% sequence identity to a recently reported Xenopus homologue xLIM-3. The gene coding for mLIM-3 maps to the murine chromosome 2, most probably within the 2B band. Based on sequence characteristics, we suggest that LIM-3 belongs to a distinct subfamily of LIM-containing homeoproteins. Ontogeny studies using in situ hybridization demonstrated that mLIM-3 transcripts can be detected on embryonic day 11 (e11) in the primordium of the hypophysis. Following a maximum between e12 and e14, lower levels persisted into adulthood, where mLIM-3 was expressed primarily in the anterior and intermediate lobes of the pituitary. These results were confirmed by Northern blot analysis in adult mice which revealed a 2.4-kb pituitary mRNA transcript. mLIM-3 transcripts were also detected in pituitary cell lines such as the somatotrophs GH3 and GH4C1, the gonadotroph alpha T3-1, and the corticotroph AtT-20 cells, but not in 20 other cell lines derived from peripheral, endocrine, and neural tissues. Starting from e11, we also observed a transient expression of mLIM-3 in the ventral part of the spinal cord, pons, and medulla oblongata, reaching a maximum at e13 and from p7 onward, the expression of this transcript is no longer detectable. mLIM-3 is also expressed in the pineal gland with high levels observed at e20. These data suggest a potential role for mLIM-3 in the transcriptional regulation of certain genes during morphogenesis and/or maintenance of the differentiated state of the pituitary, motor neurons, and pineal gland.
DNA Cell Biol 1994 Dec
PMID:The mouse homeoprotein mLIM-3 is expressed early in cells derived from the neuroepithelium and persists in adult pituitary. 781 83

The S-receptor kinase (SRK) gene which is implicated in the self-incompatibility system of Brassica oleracea is one member of a large and complex family of similar sequences. Genomic and cDNA clones were isolated for the authentic, S-linked SRK29 gene and its DNA sequence determined. Reverse transcriptase PCR (RT-PCR) was used to detect the expression of SRK29 and other members of the family in stigma, leaf and root tissues. The SRK was found to be stigma-specific whereas, for instance, K3 transcripts appeared in all three tissues. The RT-PCR analysis also demonstrated the existence of partially processed intermediates for several of the kinase transcripts and, in the case of SRK29, a product apparently resulting from the splicing of a cryptic intron. RFLP analysis of an F2 family segregating for the S29 allele was used to show S-linkage for the SRK and possibly for the K2 sequence. The K8 kinase probe also revealed a minor RFLP which segregated with the S-locus.
Plant J 1994 Dec
PMID:Expression of the S-locus receptor kinase multigene family in Brassica oleracea. 784 54


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