[PDF] Top 20 The Acidic Domain of Hepatitis C Virus NS4A Contributes to RNA Replication and Virus Particle Assembly

... Hepatitis C virus NS3-4A is a membrane-bound enzyme complex that exhibits serine protease, RNA helicase, and RNA-stimulated ATPase ...genome replication and has been recently ... See full document

... encephalomyocarditis virus (ECMV) internal ribosomal entry site (IRES) between E2-p7 and NS2-NS3 ...HCV RNA was quantified by quantitative reverse transcription (qRT)-PCR as previously described ...a ... See full document

... genome replication, whereas D333 and Y334 (JFH-1 D329 and Y330) are not ...in NS4A) but only displays a partial defect in the NS5A mutant S2201I replicon (Tellinghuisen et ...the C-terminal 30 ... See full document

... in virus assembly has not been suspected previously for HCV, several studies have suggested that the related NS3 proteins of classical flaviviruses, including both KUN and YFV (13, 20, 27), as well as ... See full document

... mutant NS4A proteins. Since NS4A is an important cofactor for HCV serine protease activity, we wondered whether the replication defects of our mutant proteins could simply be due to defective ... See full document

... viral RNA polymerase, nonstructural proteins 2B (NS2B), NS3, NS4A, and NS4B, and double-stranded RNA (dsRNA), the presumed replication intermediate, strongly suggesting that VPs are the site ... See full document

... NS4A-stimulated RNA binding and unwinding by NS3 are essential for replicon ...of NS4A function was complemented by functional genetic studies with the replicon system ...the acidic ... See full document

... gomerization, RNA-binding ability, and nuclear transport (8, 38, 41, 42, 65, ...central RNA-binding domain of HDAg (41) essential for the packag- ing and transactivation functions of the small HDAg ... See full document

... Hepatitis C virus (HCV) NS4A is a single-pass transmembrane (TM) protein essential for viral replication and particle ...the NS4A TM domain is highly conserved, ... See full document

... with RNA-transfected cells indicated that HCV proteins were detectable by 48 h posttransfection and that by this time point, viral genomes that produced in- fectious particles gave rise to higher levels of HCV ... See full document

... enhance RNA replication synergistically ...replicon, RNA replication could be mea- sured by determining the number of G418-resistant colonies obtained after selection with ...replicon, ... See full document

... Determination of adaptive mutations in the HCV coding sequence. Since adaptive mutations should be generated early during selection and become fixed in all RNA progeny, we focused our further analyses on the ... See full document

... the virus and can be used for evaluating drugs for antiviral activity or inhibition of HCV infection ...HCV replication in serum infected liver cell lines which mimics the naturally occurring HCV virions ... See full document

... on replication, attempts were made to generate more conservative mutants, with the expectation that an intermedi- ate phenotype would be ...rescue replication, and that was an equal substi- tution of ... See full document

... be the main site for adaptation by Blight et al. (7). Although they demonstrate a lack of correlation between hyperphospho- rylation and adaptation, and this was also found by our group (N. Krieger and R. Bartenschlager, ... See full document

... JFH1 replication and NS5A ...Methods. Replication capaci- ties were determined from luciferase activities approximately 96 h after trans- fection and are plotted as replication windows (relative ... See full document

... and NS4A in SGR-N cells by immunofluorescence staining ...with NS4A in the area, sharing a dotlike ...and NS4A coexist, we used double-labeling immu- noelectron microscopy on SGR-N cells using ... See full document

... of RNA interference efficiency, and negative controls were a nontar- geting siRNA pool and an siRNA pool targeting CD63, an- other tetraspanin not known to be involved in HCV ... See full document

... Hepatitis C virus (HCV) infection is a major cause of chronic hepatitis and cirrhosis and may lead to hepatocellular ...enveloped RNA virus belong- ing to the genus Hepacivirus ... See full document

... increase replication efficiency ...the replication efficiency (15; Blight and Rice, unpublished re- sults), engineering these NS3 changes into SG-5⬘HE (S2204I) did not enhance replication in our ... See full document