Abstract:
:The hepatitis C virus nonstructural NS5A protein has roles in genome replication, virus assembly, and modulation of host pathways. NS5A is a phosphoprotein, and it has been proposed that differential phosphorylation could regulate the various roles of the protein. By SDS-PAGE, two forms of NS5A with distinct apparent molecular weights can be observed, referred to as basally phosphorylated and hyperphosphorylated species. However, the sites of phosphorylation on these two species have not been unequivocally identified, hampering our understanding of the function and regulation of NS5A. To address this, we purified tagged NS5A from cells harboring a replicating subgenomic replicon and analyzed the purified protein by mass spectrometry. We identified six peptide fragments containing 12 phosphorylated residues and were able to assign four of these to serines 146, 222, and 225 and threonine 348. A serine-rich peptide fragment spanning residues 221 to 240 was highly phosphorylated. Using mutagenesis, we identified roles for a subset of these phosphoacceptors in virus genome replication. We further showed that phosphorylation at S146 regulates hyperphosphorylation, and by generating a phospho-specific antibody targeted to pS222, we identified that S222 phosphorylation predominates in the hyperphosphorylated species. Last, by introducing phosphomimetic mutations across residues 221 to 240, we demonstrated changes in the mobility of the basally phosphorylated species suggestive of a sequential phosphorylation cascade within this serine-rich cluster. We propose that this regulation could drive a conformational switch between the dimeric structures of NS5A and could also explain the different functions of the protein in the virus life cycle.
journal_name
J Viroljournal_title
Journal of virologyauthors
Ross-Thriepland D,Harris Mdoi
10.1128/JVI.03017-13subject
Has Abstractpub_date
2014-02-01 00:00:00pages
1421-32issue
3eissn
0022-538Xissn
1098-5514pii
JVI.03017-13journal_volume
88pub_type
杂志文章abstract::Type 1 human immunodeficiency viruses encoding mutated nef reading frames are 10- to 30-fold less infectious than are isogenic viruses in which the nef gene is intact. This defect in infectivity causes nef-negative viruses to grow at an attenuated rate in vitro. To investigate the mechanism of Nef-mediated enhancement...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.7.4283-4290.1996
更新日期:1996-07-01 00:00:00
abstract::We investigated how the broadly neutralizing monoclonal antibody 2F5 affects the human immunodeficiency virus type 1 envelope glycoprotein as it undergoes receptor-induced conformational changes and show that 2F5 binds both native and fusion-intermediate conformations, suggesting inhibition of a late step in virus ent...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.78.5.2627-2631.2004
更新日期:2004-03-01 00:00:00
abstract::A soluble system was developed that could support DNA replication in simian virus 40 (SV40) chromosomes. DNA synthesis in this system required the presence of purified SV40 large tumor antigen, SV40 chromosomes prepared from virus-infected monkey cells, a crude extract from HeLa cells, and several low-molecular-weight...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.10.4820-4829.1990
更新日期:1990-10-01 00:00:00
abstract::Adult T-cell leukemia (ATL) is a fatal malignancy of CD4+ T cells infected with human T-cell leukemia virus type 1 (HTLV-1). ATL cells often exhibit random gross chromosomal rearrangements that are associated with the induction and improper repair of double-stranded DNA breaks (DSBs). The viral oncoprotein Tax has bee...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00672-18
更新日期:2018-07-17 00:00:00
abstract::We examined the 5' ends of Hantaan virus (HTN) genomes and mRNAs to gain insight into the manner in which these chains were initiated. Like those of all members of the family Bunyaviridae described so far, the HTN mRNAs contained 5' terminal extensions that were heterogeneous in both length and sequence, presumably be...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.9.5754-5762.1995
更新日期:1995-09-01 00:00:00
abstract::Temperature-sensitive (ts) mutants of the T4 phage rII gene were islated and used in temperature shift experiments that revelaed two different expressions for the normal rII (rII+) gene function in vivo: (i) an early expression (0 to 12 min postinfection at 30 C) that prevents restriction of T4 growth in Escherichia c...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.16.4.974-981.1975
更新日期:1975-10-01 00:00:00
abstract::Duck embryo fibroblast (DEF) and chicken embryo fibroblast (CEF) cultures infected with Marek's disease virus were studied by combined fluorescent antibody and electron microscopy techniques. In both DEF and CEF cultures, cells containing immunofluorescent (IF) antigen also contained herpesvirus particles; conversely,...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.5.1.79-90.1970
更新日期:1970-01-01 00:00:00
abstract::We have found that the replicative helicase E1 of bovine papillomavirus type 1 (BPV-1) interacts with a key cell cycle regulator of S phase, the cyclin E-Cdk2 kinase. The E1 helicase, which interacts with cyclin E and not with Cdk2, presents the highest affinity for catalytically active kinase complexes. In addition, ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.9.7255-7262.1998
更新日期:1998-09-01 00:00:00
abstract::Murine models have suggested that CD8+ T-cell responses peak early in acute viral infections and are not sustained, but no evidence for humans has been available. To address this, we longitudinally analyzed the CD8+ T-cell response to human parvovirus B19 in acutely infected individuals. We observed striking CD8+ T-ce...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.18.12117-12121.2005
更新日期:2005-09-01 00:00:00
abstract::The human immunodeficiency virus type 1 Tat protein binds to an RNA stem-loop structure called TAR which is present at the 5' end of all human immunodeficiency virus type 1 transcripts. This binding is centered on a bulge within the stem of TAR and is an essential step in the trans-activation process which results in ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.65.10.5196-5202.1991
更新日期:1991-10-01 00:00:00
abstract::We sequenced the genes coding for the two neutralization proteins, VP4 and VP7, of human rotavirus strains L26 and L27 with subgroup I specificity but the long RNA pattern. The deduced VP7 amino acid sequence of strains L26 and L27 showed a low homology (73.6 to 81.9%) to those of rotavirus strains of the established ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.64.11.5640-5644.1990
更新日期:1990-11-01 00:00:00
abstract::Chikungunya virus (CHIKV) is a mosquito-transmitted alphavirus that has been responsible for an epidemic outbreak of unprecedented magnitude in recent years. Since then, significant efforts have been made to better understand the biology of this virus, but we still have poor knowledge of CHIKV interactions with host c...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.06390-11
更新日期:2012-03-01 00:00:00
abstract::Human influenza viruses are more efficiently isolated by inoculating patient samples into the amniotic rather than the allantoic cavity of embryonated chicken eggs. This type of cultivation selects virus variants with mutations around the hemagglutinin (HA) receptor binding site. To understand the molecular basis of t...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.71.4.3357-3362.1997
更新日期:1997-04-01 00:00:00
abstract::Hepatitis C virus (HCV) exploits cellular proteins to facilitate viral propagation. To identify the cellular factors involved in the HCV life cycle, we previously performed protein microarray assays using either HCV nonstructural 5A (NS5A) protein or core protein as a probe. Interestingly, cellular cortactin strongly ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01306-20
更新日期:2020-09-15 00:00:00
abstract::We found that recircularized high-risk (type 16 and 18) and low-risk mucosal (type 6b and 11) and cutaneous (type 5 and 8) human papillomavirus (HPV) genomes replicate readily when delivered into U2OS cells by electroporation. The replication efficiency is dependent on the amount of input HPV DNA and can be followed f...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01985-10
更新日期:2011-04-01 00:00:00
abstract::Nine pre-early polypeptides have been detected after infection with bacteriophage T5 and 10 pre-early polypeptides have been detected after infection with bacteriophage BF23. Only about one-half of the coding capacity of the redundant ends of the phage DNA, which code for pre-early proteins, is needed for these 9 to 1...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.22.2.480-488.1977
更新日期:1977-05-01 00:00:00
abstract::With the aid of three monoclonal antibodies (MAbs), a glycoprotein specifically localized to the outer envelope of vaccinia virus was shown to be encoded by the A33R gene. These MAbs reacted with a glycosylated protein that migrated as 23- to 28-kDa and 55-kDa species under reducing and nonreducing conditions, respect...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.6.3753-3762.1996
更新日期:1996-06-01 00:00:00
abstract::Although hepatitis A virus (HAV) is typically transmitted by the fecal-oral route, little is known of its interactions with cells of the gastrointestinal tract. We studied the replication of HAV in polarized cultures of Caco-2 cells, a human cell line which retains many differentiated functions of small intestinal epi...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.14.6476-6484.2000
更新日期:2000-07-01 00:00:00
abstract::We have studied the unintegrated infectious DNA of Harvey sarcoma virus (Ha-SV) and Moloney leukemia virus (Mo-MuLV). The source of infectious viral DNA was the Hirt supernatant fraction from cells acutely infected with Ha-SV and Mo-MuLV. To obtain a direct quantitative assay for infectious viral DNA, recipient mouse ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.26.2.291-298.1978
更新日期:1978-05-01 00:00:00
abstract::Coevolution of virus and host is a process that emerges in persistent virus infections. Here we studied the coevolutionary development of coxsackievirus B3 (CVB3) and cardiac myocytes representing the major target cells of CVB3 in the heart in a newly established persistently CVB3-infected murine cardiac myocyte cell ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00621-11
更新日期:2011-12-01 00:00:00
abstract::Rigid amphipathic fusion inhibitors (RAFIs) are lipophilic inverted-cone-shaped molecules thought to antagonize the membrane curvature transitions that occur during virus-cell fusion and are broad-spectrum antivirals against enveloped viruses (Broad-SAVE). Here, we show that RAFIs act like membrane-binding photosensit...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02907-13
更新日期:2014-02-01 00:00:00
abstract::The association between the level of class I major histocompatibility (MHC) antigen expression and the tumorigenic phenotype was determined for cells from a series of 15 lines of adenovirus type 2 (Ad2)-, Ad12-, and simian virus 40 (SV40)-transformed hamster cells and 16 lines of cells established from hamster tumors ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.62.8.2755-2761.1988
更新日期:1988-08-01 00:00:00
abstract::The architecture of a retroviral genome presents some unusual features for transcriptional regulation because of duplication of the transcriptional control sequences in the 5' and 3' long terminal repeats (LTRs). We have studied the transcriptional activity of the 5' and 3' LTRs of human immunodeficiency virus type 1 ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.68.6.3830-3840.1994
更新日期:1994-06-01 00:00:00
abstract::Persistent human immunodeficiency virus (HIV) infection of human monocytes and macrophages increases I kappa B alpha degradation, resulting in the activation of NF-kappa B, a key transcription factor in the regulation of the HIV long terminal repeat. The signal transduction pathways leading to NF-kappa B activation in...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.4.2332-2338.1996
更新日期:1996-04-01 00:00:00
abstract::The hemagglutinin-neuraminidase (HN) glycoprotein plays a critical role in parainfluenza virus replication. We recently found that in addition to the catalytic binding site, HN of human parainfluenza virus type 1 (hPIV-1) may have a second receptor-binding site covered by an N-linked glycan at residue 173, which is ne...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02331-09
更新日期:2010-03-01 00:00:00
abstract::S-Adenosylhomocysteine was found to have no effect on Epstein-Barr virus-induced transformation of B-lymphocytes and to stimulate viral capsid antigen expression only slightly in the FF41-1 cell line. In contrast, the S-adenosylhomocysteine analogs sinefungin and S-isobutyladenosine inhibited Epstein-Barr virus transf...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.61.1.221-224.1987
更新日期:1987-01-01 00:00:00
abstract::A new group of nucleocytoplasmic shuttling proteins has recently been identified in the structural proteins encoded by several alphaherpesvirus UL47 genes. Nuclear import and export signals for the bovine herpesvirus type 1 UL47 protein (VP8 or bUL47) have been described previously. Here, we study the trafficking of b...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01322-06
更新日期:2006-10-01 00:00:00
abstract::The protein product of varicella-zoster virus (VZV) ORF47 is a serine/threonine protein kinase and tegument component. Evaluation of two recombinants of the Oka strain, rOka47DeltaC, with a C-terminal truncation of ORF47, and rOka47D-N, with a point mutation in the conserved kinase motif, showed that ORF47 kinase func...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.78.23.13293-13305.2004
更新日期:2004-12-01 00:00:00
abstract::Murine norovirus (MNV), a prevalent pathogen of laboratory mice, shares many characteristics with human noroviruses. Previous results indicated that passage of MNV1 in the macrophage cell line RAW 264.7 results in attenuation in STAT1-deficient mice (C. E. Wobus, S. M. Karst, L. B. Thackray, K. O. Chang, S. V. Sosnovt...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00237-08
更新日期:2008-08-01 00:00:00
abstract::Mitochondrial DNA synthesis in adenovirus type 2-infected HeLa cells was measured at various times from 0 to 24 h postinfection. Although viral infection effectively turned off host chromosomal DNA synthesis, mitochondrial DNA synthesis was not inhibited. These findings indicate a dissociation between the regulation o...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.22.2.340-345.1977
更新日期:1977-05-01 00:00:00