Abstract:
:Previously, the cyclophilin inhibitors cyclosporine (CsA) and alisporivir (ALV) were shown to inhibit the replication of diverse RNA viruses, including arteriviruses and coronaviruses, which both belong to the order Nidovirales In this study, we aimed to identify arterivirus proteins involved in the mode of action of cyclophilin inhibitors and to investigate how these compounds inhibit arterivirus RNA synthesis in the infected cell. Repeated passaging of the arterivirus prototype equine arteritis virus (EAV) in the presence of CsA revealed that reduced drug sensitivity is associated with the emergence of adaptive mutations in nonstructural protein 5 (nsp5), one of the transmembrane subunits of the arterivirus replicase polyprotein. Introduction of singular nsp5 mutations (nsp5 Q21R, Y113H, or A134V) led to an ∼2-fold decrease in sensitivity to CsA treatment, whereas combinations of mutations further increased EAV's CsA resistance. The detailed experimental characterization of engineered EAV mutants harboring CsA resistance mutations implicated nsp5 in arterivirus RNA synthesis. Particularly, in an in vitro assay, EAV RNA synthesis was far less sensitive to CsA treatment when nsp5 contained the adaptive mutations mentioned above. Interestingly, for increased sensitivity to the closely related drug ALV, CsA-resistant nsp5 mutants required the incorporation of an additional adaptive mutation, which resided in nsp2 (H114R), another transmembrane subunit of the arterivirus replicase. Our study provides the first evidence for the involvement of nsp2 and nsp5 in the mechanism underlying the inhibition of arterivirus replication by cyclophilin inhibitors.IMPORTANCE Currently, no approved treatments are available to combat infections with nidoviruses, a group of positive-stranded RNA viruses, including important zoonotic and veterinary pathogens. Previously, the cyclophilin inhibitors cyclosporine (CsA) and alisporivir (ALV) were shown to inhibit the replication of diverse nidoviruses (both arteriviruses and coronaviruses), and they may thus represent a class of pan-nidovirus inhibitors. In this study, using the arterivirus prototype equine arteritis virus, we have established that resistance to CsA and ALV treatment is associated with adaptive mutations in two transmembrane subunits of the viral replication machinery, nonstructural proteins 2 and 5. This is the first evidence for the involvement of specific replicase subunits of arteriviruses in the mechanism underlying the inhibition of their replication by cyclophilin inhibitors. Understanding this mechanism of action is of major importance to guide future drug design, both for nidoviruses and for other RNA viruses inhibited by these compounds.
journal_name
J Viroljournal_title
Journal of virologyauthors
de Wilde AH,Boomaars-van der Zanden AL,de Jong AWM,Bárcena M,Snijder EJ,Posthuma CCdoi
10.1128/JVI.00490-19subject
Has Abstractpub_date
2019-08-28 00:00:00issue
18eissn
0022-538Xissn
1098-5514pii
JVI.00490-19journal_volume
93pub_type
杂志文章abstract::The capsid mRNA transcripts of human bocavirus 1 (HBoV1) can be generated by alternative splicing from the mRNA precursor transcribed from the P5 promoter. However, the alternative translation regulation mechanism of capsid mRNA transcripts is largely unknown. Here we report that the polycistronic capsid mRNA transcri...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00443-18
更新日期:2018-10-12 00:00:00
abstract::The pandemic threat posed by emerging zoonotic influenza A viruses necessitates development of antiviral agents effective against various antigenic subtypes. Human monoclonal antibody (hMAb) targeting the hemagglutinin (HA) stalk offers a promising approach to control influenza virus infections. Here, we investigated ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01284-16
更新日期:2016-11-14 00:00:00
abstract::Harvard Medical School convened a meeting of biomedical and clinical experts on 5 March 2015 on the topic of "Rethinking the Response to Emerging Microbes: Vaccines and Therapeutics in the Ebola Era," with the goals of discussing the lessons from the recent Ebola outbreak and using those lessons as a case study to aid...
journal_title:Journal of virology
pub_type: 共识发展会议,杂志文章
doi:10.1128/JVI.01251-15
更新日期:2015-08-01 00:00:00
abstract::The severe acute respiratory syndrome coronavirus (SARS-CoV) encodes numerous accessory proteins whose importance in the natural infection process is currently unclear. One of these accessory proteins is set apart by its function in the context of a related murine hepatitis virus (MHV) infection. SARS-CoV protein 6 in...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02406-07
更新日期:2008-07-01 00:00:00
abstract::Enhancement of cell death is a distinguishing feature of H1N1 influenza virus A/Puerto Rico/8/34 protein PB1-F2. Comparing the sequences (amino acids [aa] 61 to 87 using PB1-F2 amino acid numbering) of the PB1-F2-derived C-terminal peptides from influenza A viruses inducing high or low levels of cell death, we identif...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01373-13
更新日期:2014-01-01 00:00:00
abstract::Serological surveys have revealed that 30 to 50% of wild-caught African green monkeys have antibodies reactive to simian immunodeficiency virus (SIV), a retrovirus related to human immunodeficiency virus (HIV). Although the nucleotide sequence of one SIVagm isolate, Tyo1, was recently reported, the extent of genetic v...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.63.4.1800-1802.1989
更新日期:1989-04-01 00:00:00
abstract::Human cells infected with adenovirus type 2 (Ad2) or Ad5 require VAI RNA for efficient translation of viral mRNAs at late times after infection. The Ad5 mutant dl-sub720 synthesized neither virus-associated I (VAI) nor VAII RNAs, and infection of human cells with this mutant resulted in reduced virion polypeptide synt...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.60.2.363-368.1986
更新日期:1986-11-01 00:00:00
abstract::The Autographa californica multiple nucleopolyhedrovirus (AcMNPV) glycoprotein GP64 is the major envelope protein of the budded virus (BV). GP64 is a class III fusion protein that mediates BV attachment to the cell surface and low-pH-triggered membrane fusion between the BV envelope and the endosome membrane during en...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00813-12
更新日期:2012-09-01 00:00:00
abstract::We set out to test the hypothesis that interleukin-22 (IL-22), a cytokine crucial for epithelial cell homeostasis and recovery from tissue injury, would be protective during influenza virus infection. Recent studies have identified phenotypically and functionally unique intestinal NK cells capable of producing the cyt...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00187-10
更新日期:2010-08-01 00:00:00
abstract::Regulatory T (T(R)) cells maintain tolerance to self-antigens and control immune responses to alloantigens after organ transplantation. Here, we show that CD4(+) CD25(+) human T(R) cells suppress virus-specific T-cell responses. Depletion of T(R) cells from peripheral blood mononuclear cells enhances T-cell responses ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.78.5.2454-2459.2004
更新日期:2004-03-01 00:00:00
abstract:UNLABELLED:A hallmark of Ebola virus (EBOV) infection is the formation of viral inclusions in the cytoplasm of infected cells. These viral inclusions contain the EBOV nucleocapsids and are sites of viral replication and nucleocapsid maturation. Although there is growing evidence that viral inclusions create a protected...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00459-16
更新日期:2016-07-27 00:00:00
abstract::The RNA-dependent RNA polymerase (NS5B) of hepatitis C virus (HCV) is an unusually attractive target for drug discovery since it contains five distinct drugable sites. The success of novel antiviral therapies will require nonnucleoside inhibitors to be active in at least patients infected with HCV of subtypes 1a and 1...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01980-09
更新日期:2010-03-01 00:00:00
abstract::Rotaviruses are the leading etiological agents of acute gastroenteritis in infants and young children worldwide. These nonenveloped viruses enter cells using different types of endocytosis and, depending on the virus strain, travel to different endosomal compartments before exiting to the cytosolic space. In this Gem,...
journal_title:Journal of virology
pub_type: 杂志文章,评审
doi:10.1128/JVI.01787-14
更新日期:2015-01-15 00:00:00
abstract::Viruses display a wide range of genomic profiles and, consequently, a variety of gene expression strategies. Specific sequences associated with transcriptional processes have been described in viruses, and putative promoter motifs have been elucidated for some nucleocytoplasmic large DNA viruses (NCLDV). Among NCLDV, ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01088-17
更新日期:2017-10-13 00:00:00
abstract::The Borna disease virus (BDV) is the prototype member of the Bornaviridae, and it replicates in the cell nucleus. The BDV p24P and p40N proteins carry nuclear localization signals (NLS) and are found in the nuclei of infected cells. The BDV p10 protein does not have an NLS, but it binds with P and/or N and is transloc...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.7.3413-3417.2000
更新日期:2000-04-01 00:00:00
abstract::Live attenuated RNA viruses make highly efficient vaccines. Among them, measles virus (MV) vaccine has been given to a very large number of children and has been shown to be highly efficacious and safe. Therefore, this vaccine might be a very promising vector to immunize children against both measles and other infecti...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.77.21.11546-11554.2003
更新日期:2003-11-01 00:00:00
abstract::To identify the domains of Vpr that are involved nuclear localization, we transfected HeLa cells with a panel of expression vectors that encode mutant Vpr protein with deletions or substitutions within putative domains. Immunofluorescence staining of transfected cells revealed that wild-type Vpr was localized predomin...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.15.7179-7186.2000
更新日期:2000-08-01 00:00:00
abstract:UNLABELLED:Human metapneumovirus (HMPV), a recently discovered paramyxovirus, infects nearly 100% of the world population and causes severe respiratory disease in infants, the elderly, and immunocompromised patients. We previously showed that HMPV binds heparan sulfate proteoglycans (HSPGs) and that HMPV binding requir...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01362-16
更新日期:2016-09-29 00:00:00
abstract:UNLABELLED:Marek's disease virus (MDV), an alphaherpesvirus, is the causative agent of a lethal disease in chickens characterized by generalized nerve inflammation and rapid lymphoma development. The extensive colinearity of the MDV genome with those of related herpesviruses has eased functional characterization of man...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02687-14
更新日期:2015-01-15 00:00:00
abstract::The bluetongue virus (BTV) minor protein VP4, with molecular mass of 76 kDa, is one of the seven structural proteins and is located within the inner capsid of the virion. The protein has a putative leucine zipper near the carboxy terminus of the protein. In this study, we have investigated the functional activity of t...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.72.4.2983-2990.1998
更新日期:1998-04-01 00:00:00
abstract::Tryptic digests of four polypeptides found in Kunjin virus-infected Vero cells, NV5, NV4, V3, and NV3, were compared by peptide mapping. The polypeptides to be analyzed were labeled with radioactive methionine and separated by electrophoresis through polyacrylamide gels containing sodium dodecyl sulfate. Because infec...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.24.2.651-661.1977
更新日期:1977-11-01 00:00:00
abstract::The latency-related (LR) gene of bovine herpesvirus 1 (BHV-1) is abundantly expressed during latency. A mutant BHV-1 strain that contains three stop codons at the 5' terminus of the LR gene (LR mutant) does not reactivate from latency. This study demonstrates that the LR mutant does not express open reading frame 2 or...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.78.6.3184-3189.2004
更新日期:2004-03-01 00:00:00
abstract::Human herpesvirus 8 (HHV-8) viral interleukin-6 (vIL-6) is a cytokine that is poorly secreted and localized largely to the endoplasmic reticulum (ER). It has been implicated, along with other HHV-8 proinflammatory and/or angiogenic viral proteins, in HHV-8-associated Kaposi's sarcoma, primary effusion lymphoma (PEL), ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00909-20
更新日期:2020-09-15 00:00:00
abstract::Simian virus 40 large T antigen (large T) in the early and the late phases of infection differs significantly in its sequence-specific DNA-binding and ATPase activities, indicating that different large-T populations participate in virus-specific events at various stages of the infectious cycle. To further characterize...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.63.5.2308-2316.1989
更新日期:1989-05-01 00:00:00
abstract::The cloned murine cytolytic T-lymphocyte line IE1-IL and several sublines detect a murine cytomegalovirus immediate-early (IE) membrane determinant in conjunction with Ld class I major histocompatibility glycoprotein. The lines retained cytolytic activity, strict antigen specificity, and self-restriction even when ada...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.60.3.1125-1129.1986
更新日期:1986-12-01 00:00:00
abstract::Pseudorabies virus (PRV), a member of the Alphaherpesvirinae, has a complex multilayered extracellular virion that is structurally conserved among other herpesviruses. PRV virions contain a double-stranded DNA genome within a proteinaceous capsid surrounded by the tegument, a layer of viral and cellular proteins. The ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02253-10
更新日期:2011-07-01 00:00:00
abstract::During the S phase of the eukaryotic cell cycle and in virus-infected cells, DNA replication takes place at discrete sites in the nucleus, although it is not clear how the proteins involved in the replicative process are directed to these sites. Nuclear factor I is a cellular, sequence-specific DNA-binding protein uti...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.66.5.3140-3150.1992
更新日期:1992-05-01 00:00:00
abstract::The basis for the switch from CCR5 to CXCR4 coreceptor usage seen in approximately 50% of human immunodeficiency virus type 1 (HIV-1) subtype B-infected individuals as disease advances is not well understood. Among the reasons proposed are target cell limitation and better immune recognition of the CXCR4 (X4)-tropic c...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00759-07
更新日期:2007-08-01 00:00:00
abstract::The infected cell polypeptide 4 (ICP4) of herpes simplex virus 1 (HSV-1) is a regulator of viral transcription that is required for productive infection. Since viral genes are transcribed by cellular RNA polymerase II (RNA pol II), ICP4 must interact with components of the pol II machinery to regulate viral gene expre...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00385-11
更新日期:2011-06-01 00:00:00
abstract::Human cytomegalovirus (HCMV) virions are composed of a DNA-containing nucleocapsid surrounded by a tegument layer and host-derived lipid envelope studded with virally encoded glycoproteins. These complex virions are estimated to be composed of more than 50 viral proteins. Assembly of HCMV virions is poorly understood,...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01551-10
更新日期:2011-01-01 00:00:00