Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “DsRNA”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 145 records · Page 8Linked to original sources
PubMed1980

Interferon induction by viruses. VI. Reovirus: virion genome dsRNA as the interferon inducer in aged chick embryo cells.

The interferon-inducing particle (IFP) activity of avian and human reoviruses in aged chick embryo cells was determined by analyzing dose (multiplicity)-response (interferon yield) curves. These curves fit best a model in which each cell infected with greater than or equal to 1 IFP produces a quantum yield of interferon. Avian reovirus stocks contained as many as 60 times more IFP than plaque-forming particles (PFP). Upon UV-irradiation the ratio of IFP:PFP became 197, suggesting that virtually every physical particle of avian reovirus could function as an interferon-inducing particle. Thus, about one-third of the non-infectious particles were intrinsically IFP and the other two-thirds could be converted to IFP status at an optimal dose of UV radiation, the equivalent of 9.4 lethal hits, i.e., 8000 ergs/mm2. UV-irradiated avian reovirus induced about twice the usual yield of interferon on a per cell basis. Wildtype human reovirus (type 3) and mutants ts201(A,RNA+) and ts447(C,RNA-) were excellent inducers of interferon, but only about 1 in 3 infectious particles functioned as an interferon-inducing particle, meaning that virtually all physical particles failed to function as IFP, UV-irradiation of human reoviruses resulted in a slight loss of IFP activity. Our data support the hypothesis that virion genome dsRNA constitutes the interferon inducer moiety of avian reoviruses and that in its permissive host cell the processing of genome dsRNA from most particles to a putative recognition site in the cytoplasm occurs naturally with a high probability. For human reovirus this is a much rarer event which may be intrinsic only to infectious virus, and may require limited transcription for expression. The sensitivity of the avian reovirus-aged chick embryo cell system recommends it for further study on the mechanism of interferon induction by virions containing pre-existing dsRNA.

Animals↗
PubMed2000

The polymerase subunit of a dsRNA virus plays a central role in the regulation of viral RNA metabolism.

Bacteriophage φ6 has a three-segmented double-stranded (ds) RNA genome, which resides inside a polymerase complex particle throughout the entire life cycle of the virus. The polymerase subunit P2, a minor constituent of the polymerase complex, has previously been reported to replicate both φ6-specific and heterologous single-stranded (ss) RNAs, giving rise to dsRNA products. In this study, we show that the enzyme is also able to use dsRNA templates to perform semi-conservative RNA transcription in vitro without the assistance of other proteins. The polymerase synthesizes predominantly plus-sense copies of φ6 dsRNA, medium and small segments being more efficient templates than the large one. This distribution of the test-tube reaction products faithfully mimics viral transcription in vivo. Experiments with chimeric ssRNAs and dsRNAs show that short terminal nucleotide sequences can account for the difference in efficiency of RNA synthesis. Taken together, these results suggest a model explaining important aspects of viral RNA metabolism regulation in terms of enzymatic properties of the polymerase subunit.

Bacteriophage phi 6↗
PubMed1989

Activation of interferon-regulated, dsRNA-dependent enzymes by human immunodeficiency virus-1 leader RNA.

Human immunodeficiency virus-1 (HIV-1) leader RNA, which contains double-stranded regions due to inverted repeats, was shown to activate the dsRNA-dependent enzymes associated with the interferon system. HIV-1 leader RNA produced in vitro using SP6 RNA polymerase was characterized using probes for antisense and sense-strand RNA. The RNA preparation was free from significant levels of antisense RNA. HIV-1 leader RNA was shown to activate dsRNA-dependent protein kinase in a cell-free system from interferon-treated HeLa cells. Affinity resins, consisting of HIV-1 leader RNA covalently attached to cellulose, immobilized and activated dsRNA-dependent protein kinase and 2-5A-synthetase. HIV-1 leader RNA, therefore, may be a contributing factor in the mechanism by which interferon inhibits HIV replication.

2',5'-Oligoadenylate Synthetase↗
PubMed2001

Apoptosis is promoted by the dsRNA-activated factor (DRAF1) during viral infection independent of the action of interferon or p53.

An apoptotic cellular defense mechanism is triggered in response to viral dsRNA generated during the course of infection by many DNA and RNA viruses. We demonstrate that apoptosis induced by dsRNA or a paramyxovirus is independent of the action of interferon as it can proceed in a variety of cell lines and primary cells deficient in an interferon response. Initiation of apoptosis appears to be triggered by activation of a cellular transcription factor, the dsRNA-activated factor (DRAF1). DRAF1 is composed of interferon regulatory factor 3 (IRF-3) and the transcriptional coactivators CREB binding protein (CBP) or p300. We find that activation of IRF-3 in the absence of viral infection stimulates apoptosis. In addition, a negative interfering mutant blocks both target gene induction and apoptosis, demonstrating a requirement for gene expression by IRF-3/DRAF1 to promote apoptosis. IRF-3/DRAF1 target gene expression is also induced in response to a distinct apoptotic stimulus, the DNA damaging agent etoposide. The activity of the p53 tumor suppressor does not appear to be required for IRF-3/DRAF1-mediated apoptosis.

Apoptosis↗
PubMed1992

Binding of influenza A virus NS1 protein to dsRNA in vitro.

The non-structural protein NS1 of influenza A virus exhibits two modes of RNA-binding activity. One is sequence-specific binding to minus-sense virus RNA with either a 5'- or 3'-terminal common sequence as reported previously. The other was identified as binding to dsRNA and this activity did not show sequence specificity. The affinity of binding to dsRNA was much higher than that to ssRNA. A short miniature virion RNA forming a panhandle structure by pairing between the 5'- and 3'-terminal common sequences bound NS1 with higher affinity and stability than did a dsRNA of similar sequence and length.

Base Sequence↗
PubMed1995

Cherry virus A: cDNA cloning of dsRNA, nucleotide sequence analysis and serology reveal a new plant capillovirus in sweet cherry.

The nucleotide sequence (7383 nucleotides) of a newly identified member of the genus Capillovirus, cherry virus A (CVA), was obtained from cDNA clones. The cDNA was generated from dsRNA extracted from plant tissue infected with little cherry virus (LCV). Small amounts of LCV dsRNA served as template nucleic acid and enabled the construction of a library of which, unexpectedly, 7.5% of the recombinant plasmids were specific for CVA. The genome organization of CVA resembles that of apple stem grooving virus (ASGV), the type member of the genus Capillovirus and is composed of a 266 kDa polyprotein (ORF1), a 52 kDa ORF2 located within ORF1 and a poly(A) tail. The 266 kDa ORF1 contains all the elements of a replication-related protein and has high identity with 'Sindbis-like' viruses. The ORF encodes the coat protein (CP) in the C-terminal region. The 52 kDa ORF2 has high identities with the putative viral cell-to-cell movement proteins of capillo- and trichoviruses. The CP was identified in immunoblot analysis and estimated to have a molecular mass of 24 kDa. Antiserum was obtained by expression of antigens as fusion proteins in Escherichia coli. There is significant sequence identity between CVA CP and the corresponding proteins of other capillo- and trichoviruses. However, no serological cross-reaction was obtained in immunoblot analysis with ASGV, apple chlorotic leafspot trichovirus (ACLSV), apple stem pitting virus (ASPV) and cherry mottle leaf virus (CMLV) antisera. Flexuous filamentous CVA virions were identified in extracts of sweet cherry by immunosorbent electron microscopy (ISEM) and decorated with the antiserum to the fusion protein. CVA was identified in three cherry sources of different disease status by ISEM, immunoblot analysis and hybridization to dsRNA. CVA is not closely related to any of the currently described diseases in cherry but it has all the properties of a capillovirus. It is suggested that CVA should be classified as a new member of the genus capillovirus.

Amino Acid Sequence↗
PubMed1996

Interference of reovirus strains occurs between the stages of uncoating and dsRNA accumulation.

Interference of wild-type reovirus growth by some temperature-sensitive (ts) mutant viruses under non-permissive conditions or by other wild-type isolates has been demonstrated; however, the stage of the virus replication cycle at which interference occurs has not been defined. Examination of the time-course of the yields of T1 Lang (T1L) dsRNA in the progeny of mixed infections of T1L with T3 Dearing (T3D) or with a panel of T3D ts mutants at a non-permissive temperature revealed that interference takes place by 8-10 h post-infection and occurs prior to or at the same time as accumulation of reovirus dsRNA. Taken together with our previous results, these data indicate that interference occurs during a window between virus uncoating and synthesis of dsRNA in the reovirus replication cycle, probably at the stage of assembly of primary reovirus particles.

Capsid↗
PubMed2003

Transport of dsRNA into cells by the transmembrane protein SID-1.

RNA interference (RNAi) spreads systemically in plants and nematodes to silence gene expression distant from the site of initiation. We previously identified a gene, sid-1, essential for systemic but not cell-autonomous RNAi in Caenorhabditis elegans. Here, we demonstrate that SID-1 is a multispan transmembrane protein that sensitizes Drosophila cells to soaking RNAi with a potency that is dependent on double-stranded RNA (dsRNA) length. Further analyses revealed that SID-1 enables passive cellular uptake of dsRNA. These data indicate that systemic RNAi in C. elegans involves SID-1-mediated intercellular transport of dsRNA.

Adenosine Triphosphate↗
PubMed2005

Activation of dsRNA dependent protein kinase PKR in Karpas299 does not lead to cell death.

Activated double-stranded RNA (dsRNA)-dependent protein kinase PKR is a potent growth inhibitory protein that is primarily activated in virally infected cells, inducing them to die. We have recently shown that PKR can be selectively activated in cancer cells, by in situ generation of dsRNA following introduction of antisense RNA complementary to an RNA expressed specifically in the cancer cell. The feasibility of this approach was demonstrated using a glioblastoma line that overexpresses a truncated form of the EGFR. PKR and its signaling pathway are not restricted to a given cell line; therefore, in principle, this dsRNA killing approach can be applied to any cancer that expresses unique RNA sequences. Nonetheless, applying this approach to Karpas299 cells, from a T-cell non-Hodgkin's lymphoma that harbors the NPM/ALK translocation, did not result in cell death, implying that PKR signaling pathway is repressed in this cell line. Indeed, the phosphorylation of eIF2alpha by PKR was impaired in Karpas299 cells. Furthermore, levels of the cellular inhibitor p67 were elevated in these cells. Long antisense, as well as RNAi for p67, delivered into Karpas299 cells by adenoviruses, reduced p67 levels. The reduction in p67 levels led to increased phosphorylation of eIF2alpha, and an additive effect was achieved by coinfection with NPM/ALK-AS encoding adenoviruses. Infection with these adenoviruses, however, did not promote growth inhibition. These findings imply that anti-apoptotic mechanisms counteract PKR signaling in this T-cell non-Hodgkin's lymphoma.

Adenoviridae↗
PubMedDate not supplied

[Activity of dsRNA-dependent protein kinase in rat lymphoid cells under the effect of X-ray irradiation].

The activity of dsRNA-dependent protein kinase, which is the key enzyme of the interferon signal system, was studied in the rat spleen and thymus lymphocytes under the influence of X-ray irradiation at 0.5 and 1 Gy doses and interferon inducers administration. An increase of the enzyme activity was established in the presence of FGA, concanavaline A, poly(I).poly(C) in vitro. The effect is intensified under the irradiation by 0.5 Gy dose. The protein kinase activity in lymphocytes is amplified in proportion to poly(I).poly(C) concentration, that was most pronounced in the irradiated animals. The comparative analysis of the action of interferon inducers on the dsRNA-dependent protein kinase activity was carried out. Two biological systems were used: in vivo (when the preparations were injected to the experimental animals) and in vivo (under the preincubation of isolated lymphocytes with the inducers). It was shown that the combined action of radiation and interferon inducers causes the stimulation of dsRNA-dependent protein kinase activity.

2',5'-Oligoadenylate Synthetase↗
PubMed2005

[Molecular polymorphism of viral dsRNA of yeast Saccharomyces paradoxus].

An analysis of 53 strains of yeast Saccharomyces paradoxus (YSP) of different geographic origins enabled us, for the first time, to find viral double-stranded RNA (L and M fractions) in YSP and to study natural polymorphism. As in the cultured Scerevisiae, the size of L dsRNA was constant (4.5 kb). The size of minor M dsRNA varied from 1.5 to 2.4 k.b. In YSP, we determined 7 types of M dsRNA (M1-M7), which were not connected with the source of isolation or geographic origin of the host strains.

Asia↗
PubMed1991

[Regulation of Ca(2+)-dsRNA for proliferation and terminal differentiation processes of human fibroblasts and HeLa cells].

Ca2+ complexes of dsRNA, poly(dA) and poly(dT) of yeast low molecular weight RNA produce a pronounced mitogenic effect on human fibroblasts at early stages of fibroblast proliferation in culture. At later stages of cell cultivation Ca(2+)-dsRNA stimulates terminal differentiation by inducing the synthesis of proteins characteristic of the postmitotic population of human fibroblasts undergoing terminal differentiation. Ca(2+)-dsRNA produces a stimulating effect on c-fos and c-jun gene transcription in fibroblasts and HeLa-S-3.

Calcium↗
PubMedDate not supplied

[The effect of high molecular weight interferon inducers (dsRNA) on virus adsorption].

Interrelationship between phosphorylation of plasmatic membrane proteins in brain cells and the rate of mice encephalomyocarditis virus adsorption was studied. Phosphorylation of proteins induced by dsRNA (laryphane) was most distinctly manifested in membrane fraction and cytosol of rat brain neuronal cells. Similarity of molecular mass spectra in dsRNA- and cAMP-dependent phosphorylation enabled to suggest that dsRNA activated protein kinase. An increase in the rate of plasmatic membrane proteins phosphorylation appears to correlate with inhibition of virus adsorption as shown by studies of these reactions dynamics. Under conditions of considerable increase in the rate of membrane proteins phosphorylation adsorption of virus on cells was distinctly inhibited.

Adsorption↗
PubMed1988

[Immunomodulating action of a preparation of yeast dsRNA on cellular immunity in mice].

The effect of a dsRNA preparation (an interferon inductor) on DTH induced by SRBC was studied. It was shown that at optimal antigenic load the dsRNA preparation inhibited DTH whereas at suboptimal and supraoptimal loads the preparation stimulated it. The findings indicated that the dsRNA preparation had an immunoregulatory effect. The immunoregulatory properties of the preparation must be associated with its action on the lymphocyte suppressor cells and macrophages.

Adjuvants, Immunologic↗
PubMed1997

Analysis of Xenopus dsRNA adenosine deaminase cDNAs reveals similarities to DNA methyltransferases.

We isolated two similar, but distinct, cDNA classes that encode Xenopus double-stranded RNA (dsRNA) adenosine deaminase. The longest, full-length open reading frame (ORF) predicts a 1,270-amino acid protein of 138,754 Da that is similar in size and about 50% identical to proteins encoded by mammalian cDNAs, yet larger than the 120-kDa protein purified from Xenopus eggs. Alignments of the Xenopus and mammalian ORFs show N-terminal heterogeneity, three conserved dsRNA binding motifs (dsRBMs), and strongly conserved carboxyl termini. Consistent with the observation of two cDNA classes, northern analyses of Xenopus oocyte poly A+ RNA show at least three mRNA species. Multiple nuclear polyadenylation hexamers and putative cytoplasmic polyadenylation elements were found in the 3' UTRs of cDNAs corresponding to the largest mRNA. In vitro translation experiments show that the cDNAs encode active deaminases and that the entire N-terminus and first dsRBM are dispensable for deaminase activity. Importantly, an analysis of the C-termini of five known dsRNA adenosine deaminases, and two putative deaminases, reveals motifs that are strikingly similar to the conserved motifs of the DNA-(adenine-N6alpha)-aminomethyltransferases and the DNA-(cytosine-5)-methyltransferases.

Adenosine Deaminase↗
PubMed2006

STAT1 contributes to dsRNA inhibition of liver regeneration after partial hepatectomy in mice.

Increasing evidence suggests that liver regeneration is suppressed in patients with chronic HCV infection; however, the underlying mechanisms remain unclear. Previously, we demonstrated that injection of the synthetic double-stranded RNA (dsRNA) poly I:C to mimic viral infection suppresses liver regeneration in the partial hepatectomy (PHx) model, whereby IFN-gamma contributes to the inhibition. In this study, we examined the role of the IFN-gamma-activated downstream signal (STAT1) and genes (IRF-1, p21(cip1), and SOCS1) in liver regeneration and hepatocyte proliferation. Results show that disruption of the STAT1 gene abolished poly I:C suppression of liver regeneration and the inhibitory effect of poly I:C on liver regeneration was diminished in IRF-1(-/-) and p21(cip1-/-)mice. Treatment with IFN-gamma in vitro inhibited cell proliferation of wild-type mouse hepatocytes, but not STAT1(-/-) hepatocytes. The inhibitory effect of IFN-gamma on cell proliferation was also diminished in IRF-1(-/-) and p21(cip1-/-) hepatocytes, but enhanced in SOCS1(-/-) hepatocytes. Hepatocyte proliferation was unaffected by treatment with poly I:C alone, but when hepatocytes were co-cultured with liver lymphocytes, proliferation was inhibited by IFN-gamma/STAT1-dependent mechanisms. Moreover, in HCV-infected livers with cirrhosis, activation of STAT1 was detected and correlated positively with liver injury (elevated serum levels of AST) but negatively with hepatocyte proliferation (hepatocyte PCNA and Ki-67 positive immunostaining). In conclusion, STAT1 is involved in dsRNA suppression of liver regeneration; not only does STAT1 activation contribute to liver injury, it may also block liver repair through inhibition of hepatocyte proliferation in HCV-infected patients, playing an important role in the pathogenesis of disease.

Animals↗
PubMed1995

NMR solution structure of a dsRNA binding domain from Drosophila staufen protein reveals homology to the N-terminal domain of ribosomal protein S5.

The double-stranded RNA binding domain (dsRBD) is an approximately 65 amino acid motif that is found in a variety of proteins that interact with double-stranded (ds) RNA, such as Escherichia coli RNase III and the dsRNA-dependent kinase, PKR. Drosophila staufen protein contains five copies of this motif, and the third of these binds dsRNA in vitro. Using multinuclear/multidimensional NMR methods, we have determined that staufen dsRBD3 forms a compact protein domain with an alpha-beta-beta-beta-alpha structure in which the two alpha-helices lie on one face of a three-stranded anti-parallel beta-sheet. This structure is very similar to that of the N-terminal domain of a prokaryotic ribosomal protein S5. Furthermore, the consensus derived from all known S5p family sequences shares several conserved residues with the dsRBD consensus sequence, indicating that the two domains share a common evolutionary origin. Using in vitro mutagenesis, we have identified several surface residues which are important for the RNA binding of the dsRBD, and these all lie on the same side of the domain. Two residues that are essential for RNA binding, F32 and K50, are also conserved in the S5 protein family, suggesting that the two domains interact with RNA in a similar way.

Amino Acid Sequence↗
PubMed1995

Structure of the dsRNA binding domain of E. coli RNase III.

The double-stranded RNA binding domain (dsRBD) is a approximately 70 residue motif found in a variety of modular proteins exhibiting diverse functions, yet always in association with dsRNA. We report here the structure of the dsRBD from RNase III, an enzyme present in most, perhaps all, living cells. It is involved in processing transcripts, such as rRNA precursors, by cleavage at short hairpin sequences. The RNase III protein consists of two modules, a approximately 150 residue N-terminal catalytic domain and a approximately 70 residue C-terminal recognition module, homologous with other dsRBDs. The structure of the dsRBD expressed in Escherichia coli has been investigated by homonuclear NMR techniques and solved with the aid of a novel calculation strategy. It was found to have an alpha-beta-beta-beta-alpha topology in which a three-stranded anti-parallel beta-sheet packs on one side against the two helices. Examination of 44 aligned dsRBD sequences reveals several conserved, positively charged residues. These residues map to the N-terminus of the second helix and a nearby loop, leading to a model for the possible contacts between the domain and dsRNA.

Amino Acid Sequence↗