Abstract:
:The Middle East respiratory syndrome coronavirus (MERS-CoV) nonstructural protein 16 (nsp16) is an S-adenosyl-l-methionine (SAM)-dependent 2'-O-methyltransferase (2'-O-MTase) that is thought to methylate the ribose 2'-OH of the first transcribed nucleotide (N1) of viral RNA cap structures. This 2'-O-MTase activity is regulated by nsp10. The 2'-O methylation prevents virus detection by cell innate immunity mechanisms and viral translation inhibition by the interferon-stimulated IFIT-1 protein. To unravel the regulation of nsp10/nsp16 2'-O-MTase activity, we used purified MERS-CoV nsp16 and nsp10. First, we showed that nsp16 recruited N7-methylated capped RNA and SAM. The SAM binding promotes the assembly of the enzymatically active nsp10/nsp16 complex that converted 7mGpppG (cap-0) into 7mGpppG2'Om (cap-1) RNA by 2'-OH methylation of N1 in a SAM-dependent manner. The subsequent release of SAH speeds up nsp10/nsp16 dissociation that stimulates the reaction turnover. Alanine mutagenesis and RNA binding assays allowed the identification of the nsp16 residues involved in RNA recognition forming the RNA binding groove (K46, K170, E203, D133, R38, Y47, and Y181) and the cap-0 binding site (Y30, Y132, and H174). Finally, we found that nsp10/nsp16 2'-O-MTase activity is sensitive to known MTase inhibitors, such as sinefungin and cap analogues. This characterization of the MERS-CoV 2'-O-MTase is a preliminary step toward the development of molecules to inhibit cap 2'-O methylation and to restore the host antiviral response. IMPORTANCE MERS-CoV codes for a cap 2'-O-methyltransferase that converts cap-0 into cap-1 structure in order to prevent virus detection by cell innate immunity mechanisms. We report the biochemical properties of MERS-CoV 2'O-methyltransferase, which is stimulated by nsp10 acting as an allosteric activator of the nsp16 2'-O-methyltransferase possibly through enhanced RNA binding affinity. In addition, we show that SAM promotes the formation of the active nsp10/nsp16 complex. Conversely, after cap methylation, the reaction turnover is speeded up by cap-1 RNA release and nsp10/nsp16 complex dissociation, at the low intracellular SAH concentration. These results suggest that SAM/SAH balance is a regulator of the 2'-O-methyltransferase activity and raises the possibility that SAH hydrolase inhibitors might interfere with CoV replication cycle. The enzymatic and RNA binding assays developed in this work were also used to identify nsp16 residues involved in cap-0 RNA recognition and to understand the action mode of known methyltransferase inhibitors.
journal_name
J Viroljournal_title
Journal of virologyauthors
Aouadi W,Blanjoie A,Vasseur JJ,Debart F,Canard B,Decroly Edoi
10.1128/JVI.02217-16subject
Has Abstractpub_date
2017-02-14 00:00:00issue
5eissn
0022-538Xissn
1098-5514pii
JVI.02217-16journal_volume
91pub_type
杂志文章abstract::The simian picornaviruses were isolated from various primate tissues during the development of general tissue culture methods in the 1950s to 1970s or from specimens derived from primates used in biomedical research. Twenty simian picornavirus serotypes are recognized, and all are presently classified within the Enter...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.76.3.1244-1251.2002
更新日期:2002-02-01 00:00:00
abstract::By using an isothermal multiply primed rolling-circle amplification protocol, the complete genomic DNA of a novel papillomavirus was amplified from a skin lesion biopsy of a Florida manatee (Trichechus manatus latirostris), one of the most endangered marine mammals in United States coastal waters. The nucleotide seque...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.78.22.12698-12702.2004
更新日期:2004-11-01 00:00:00
abstract::We constructed a matched set of plasmids to investigate the interactions of essential core sequences of the simian virus 40 replication origin with flanking regulatory sequences. Deletions of either T-antigen-binding region I or the 21-base-pair repeated promoter elements reduced replication to 50 to 70% of wild-type ...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.57.1.138-144.1986
更新日期:1986-01-01 00:00:00
abstract::Two poliovirus mutants were isolated that contain an oligonucleotide linker insertion in the 2C-coding region of the viral genome. One, 2C-31, has a strongly temperature-sensitive phenotype and the other, 2C-32, forms small plaques on HeLa cell monolayers at all temperatures. Both mutants have a severe temperature-sen...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.62.11.4016-4021.1988
更新日期:1988-11-01 00:00:00
abstract::A number of cDNA clones complementary to measles virus mRNA and 50S genome RNA have been generated. These clones have been mapped by restriction enzyme analysis and were subsequently sequenced by the method of Maxam and Gilbert (A. M. Maxam and W. Gilbert, Methods Enzymol. 65:499-560, 1980). Computer analysis of these...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.54.1.186-193.1985
更新日期:1985-04-01 00:00:00
abstract::Phosphorylation of the βC1 protein encoded by the betasatellite of tomato yellow leaf curl China virus (TYLCCNB-βC1) by SNF1-related protein kinase 1 (SnRK1) plays a critical role in defense of host plants against geminivirus infection in Nicotiana benthamiana However, how phosphorylation of TYLCCNB-βC1 impacts its pa...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00300-17
更新日期:2017-07-27 00:00:00
abstract::Proteasomes are critical for the processing of antigens for presentation through the major histocompatibility complex (MHC) class I pathway. HIV-1 Gag protein is a component of several experimental HIV-1 vaccines. Therefore, understanding the processing of HIV-1 Gag protein and the resulting epitope repertoire is esse...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01790-10
更新日期:2011-02-01 00:00:00
abstract::A system has been developed for generating chimeric yellow fever/Japanese encephalitis (YF/JE) viruses from cDNA templates encoding the structural proteins prM and E of JE virus within the backbone of a molecular clone of the YF17D strain. Chimeric viruses incorporating the proteins of two JE strains, SA14-14-2 (human...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.73.4.3095-3101.1999
更新日期:1999-04-01 00:00:00
abstract::Transgenic (Tg) mice expressing full-length bovine prion protein (BoPrP) serially propagate bovine spongiform encephalopathy (BSE) prions without posing a transmission barrier. These mice also posed no transmission barrier for Suffolk sheep scrapie prions, suggesting that cattle may be highly susceptible to some sheep...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.79.9.5259-5271.2005
更新日期:2005-05-01 00:00:00
abstract::Evidence is presented that the herpes simplex virus type 2 glycoprotein previously designated gF is antigenically related to herpes simplex virus type 1 gC (gC-1). An antiserum prepared against type 1 virion envelope proteins immunoprecipitated gF of type 2 (gF-2), and competition experiments revealed that the anti-gC...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.47.3.553-562.1983
更新日期:1983-09-01 00:00:00
abstract::Translation of encephalomyocarditis virus RNA in rabbit reticulocyte lysates in the presence of N-formyl-[(35)S]methionine-tRNA(f) (Met) revealed that a small polypeptide is cleaved from the N-terminus of the capsid protein precursor, preA, by virus-coded protease activity. Therefore, this N-terminal segment comprisin...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.45.1.439-441.1983
更新日期:1983-01-01 00:00:00
abstract::Co-infection of neonatal BALB/c mice with Friend virus (FV) complex (containing defective spleen focus-forming virus [SFFV] and endogenous N-tropic leukemia-inducing helper virus [LLV-F]) and B-tropic Tennant leukemia virus (TenLV) resulted in the inhibition of LLV-F by the Fv-1(b) gene and recovery of a TenLV pseudot...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.12.3.523-533.1973
更新日期:1973-09-01 00:00:00
abstract::In various positive-sense single-stranded RNA viruses, a low-fidelity viral RNA-dependent RNA polymerase (RdRp) confers attenuated phenotypes by increasing the mutation frequency. We report a negative-sense single-stranded RNA virus RdRp mutant strain with a mutator phenotype. Based on structural data of RdRp, rationa...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00834-19
更新日期:2019-10-29 00:00:00
abstract::In vivo decay rates for the individual T7 early mRNA species were determined. The physical half-lives, measured at 37 C, range from 1.1 min for gene 0.7 RNA to 4.5 min for gene 0.3 RNA. Physical half-lives, as observed after rifampin inhibition of RNA synthesis and polyacylamide electrophoresis of RNAs, are approximat...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.17.2.642-658.1976
更新日期:1976-02-01 00:00:00
abstract:UNLABELLED:Herpes simplex virus (HSV) replicates in the skin and mucous membranes, and initiates lytic or latent infections in sensory neurons. Assembly of progeny virions depends on the essential large tegument protein pUL36 of 3,164 amino acid residues that links the capsids to the tegument proteins pUL37 and VP16. O...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.03167-15
更新日期:2016-05-12 00:00:00
abstract::H9N2 avian influenza virus (AIV) has an extended host range, but the molecular basis underlying H9N2 AIV transmission to mammals remains unclear. We isolated more than 900 H9N2 AIVs in our 3-year surveillance in live bird markets in China from 2009 to 2012. Thirty-seven representative isolates were selected for furthe...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01141-16
更新日期:2016-10-14 00:00:00
abstract::The glycosylated Gag protein (gPr80) of murine leukemia viruses (MLVs) has been shown to exhibit multiple roles in facilitating retrovirus release, infection, and resistance to host-encoded retroviral restriction factors, such as APOBEC3, SERINC3, and SERINC5. One way in which gPr80 helps MLVs to escape host innate im...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.01530-17
更新日期:2018-02-26 00:00:00
abstract::Human T-cell lymphotropic virus type 1 (HTLV-1) enhances the growth of T lymphocytes, allowing the generation of T-lymphocyte cell lines. This report describes a limiting-dilution assay system which uses low input numbers of HTLV-1-producing cells for generation of T-lymphocyte cultures. The HTLV-1 transformants gener...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.69.10.6297-6303.1995
更新日期:1995-10-01 00:00:00
abstract::We have identified a novel brain-derived single-stranded-DNA-binding protein that interacts with a region of the human neurotropic JC virus enhancer designated the lytic control element (LCE). This nuclear factor, LCP-1 (for lytic control element-binding protein 1), specifically recognizes the LCE, as determined by ge...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.66.12.6885-6892.1992
更新日期:1992-12-01 00:00:00
abstract::In an earlier study on minus-strand DNA synthesis catalyzed by murine leukemia virus reverse transcriptase, we described a prominent pause site near the polypurine tract (J. Guo, W. Wu, Z. Y. Yuan, K. Post, R. J. Crouch, and J. G . Levin, Biochemistry 34:5018-5029, 1995). We now report that pausing at this site is due...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.70.10.7132-7142.1996
更新日期:1996-10-01 00:00:00
abstract::Several arenaviruses, chiefly Lassa virus (LASV) and Junin virus in West Africa and Argentina, respectively, cause hemorrhagic fever (HF) disease in humans that is associated with high morbidity and significant mortality. The investigation of antiviral strategies to combat HF arenaviruses is hampered by the requiremen...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02229-10
更新日期:2011-02-01 00:00:00
abstract::The human immunodeficiency virus type 1 (HIV-1) epidemic in Southeast Asia has been largely due to the emergence of clade E (HIV-1E). It has been suggested that HIV-1E is derived from a recombinant lineage of subtype A (HIV-1A) and subtype E, with multiple breakpoints along the E genome. We obtained complete genome se...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.74.22.10752-10765.2000
更新日期:2000-11-01 00:00:00
abstract::The product of the human cytomegalovirus (HCMV) UL71 gene is conserved throughout the herpesvirus family. During HCMV infection, protein pUL71 is required for efficient virion egress and is involved in the final steps of secondary envelopment leading to infectious viral particles. We found strong indications for oligo...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.06556-11
更新日期:2012-03-01 00:00:00
abstract::Vaccinia virus replication is inhibited by etoposide and mitoxantrone even though poxviruses do not encode the type II topoisomerases that are the specific targets of these drugs. Furthermore, one can isolate drug-resistant virus carrying mutations in the viral DNA ligase and yet the ligase is not known to exhibit sen...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.02723-07
更新日期:2008-06-01 00:00:00
abstract::Kaposi's sarcoma-associated herpesvirus (KSHV) plays a significant role in the development of Kaposi's sarcoma, primary effusion lymphoma, and some forms of multicentric Castleman's disease. The KSHV open reading frame K9 encodes the viral interferon (IFN) factor 1 (vIRF1), which downregulates IFN- and IRF-mediated tr...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/jvi.76.17.8797-8807.2002
更新日期:2002-09-01 00:00:00
abstract::The human T-cell leukemia virus type I (HTLV-I) envelope protein is synthesized as a gp61 precursor product cleaved into two mature proteins, a gp45 exterior protein and a gp20 anchoring the envelope at the cell membrane. Using N-glycosylation inhibitors and site-directed mutagenesis of the potential glycosylation sit...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.66.2.906-913.1992
更新日期:1992-02-01 00:00:00
abstract::N-terminal modifications of the chemokine RANTES bind to C-C chemokine receptor 5 (CCR5) and block human immunodeficiency virus type 1 (HIV-1) infection with greater efficacy than native RANTES. Modified RANTES compounds induce rapid CCR5 internalization and much slower receptor reexpression than native RANTES, sugges...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.75.2.661-671.2001
更新日期:2001-01-01 00:00:00
abstract::Protein P4, an early protein of double-stranded RNA bacteriophage phi 6, is a component of the virion-associated RNA polymerase complex and possesses a nucleoside triphosphate (NTP) phosphohydrolase activity. We have produced and characterized a panel of 20 P4-specific monoclonal antibodies. Epitope mapping using trun...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.67.5.2879-2886.1993
更新日期:1993-05-01 00:00:00
abstract::Human immunodeficiency virus type 2 (HIV-2) strains vary widely in their abilities to grow in Old World monkey (OWM) cells such as those of cynomolgus monkeys (CM). We evaluated eight HIV-2 isolates for their sensitivities to CM TRIM5alpha, an anti-HIV factor in OWM cells. We found that different HIV-2 isolates showed...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.00406-07
更新日期:2007-07-01 00:00:00
abstract::Deoxyribonucleic acid (DNA) synthesis was studied in poxvirus-infected cells by measuring (14)C-thymidine incorporation into viral and host cell DNA. A complete separation of the two species of DNA was achieved by combining the previously used "Dounce method" with a separation method based on different reannealing pro...
journal_title:Journal of virology
pub_type: 杂志文章
doi:10.1128/JVI.2.5.401-408.1968
更新日期:1968-05-01 00:00:00