Severe acute respiratory syndrome coronavirus evades antiviral signaling: role of nsp1 and rational design of an attenuated strain.
The severe acute respiratory syndrome (SARS) epidemic was caused by the spread of a previously unrecognized infectious agent, the SARS-associated coronavirus (SARS-CoV). Here we show that SARS-CoV could inhibit both virus- and interferon (IFN)-dependent signaling, two key steps of the antiviral response. We mapped a strong inhibitory activity to SARS-CoV nonstructural protein 1 (nsp1) and show that expression of nsp1 significantly inhibited the activation of all three virus-dependent signaling pathways. We show that expression of nsp1 significantly inhibited IFN-dependent signaling by decreasing the phosphorylation levels of STAT1 while having little effect on those of STAT2, JAK1, and TYK2. We engineered an attenuated mutant of nsp1 in SARS-CoV through reverse genetics, and the resulting mutant virus was viable and replicated as efficiently as wild-type virus in cells with a defective IFN response. However, mutant virus replication was strongly attenuated in cells with an intact IFN response. Thus, nsp1 is likely a virulence factor that contributes to pathogenicity by favoring SARS-CoV replication.
Wathelet
Marc G
MG
Department of Molecular and Cellular Physiology, University of Cincinnati College of Medicine, 231 Albert Sabin Way, Cincinnati, OH 45267-0576, USA. marc.wathelet@uc.edu
Orr
Melissa
M
Frieman
Matthew B
MB
Baric
Ralph S
RS
D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D024882:Drug Resistance, Viral; D017930:Genes, Reporter; D006801:Humans; D016899:Interferon-beta / Q000378:metabolism; D007372:Interferons / Q000378:metabolism; D008957:Models, Genetic; D011401:Promoter Regions, Genetic; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D045473:Severe acute respiratory syndrome-related coronavirus / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology*; D014709:Vero Cells; D017361:Viral Nonstructural Proteins / Q000378:metabolism*
D000998:Antiviral Agents; D017361:Viral Nonstructural Proteins; D016899:Interferon-beta; D007372:Interferons; C521592:Nsp1 protein, SARS coronavirus; D012324:RNA-Dependent RNA Polymerase
J Virol. 2004 Oct;78(19):10410-9
15367607
Lancet. 2003 Jul 26;362(9380):293-4
12892961
N Engl J Med. 2003 May 15;348(20):1967-76
12690091
Proc Natl Acad Sci U S A. 2006 Aug 22;103(34):12885-90
16912115
Cold Spring Harb Symp Quant Biol. 1998;63:609-20
10384326
J Gen Virol. 2003 Sep;84(Pt 9):2305-2315
12917450
Cytokine Growth Factor Rev. 2001 Jun-Sep;12(2-3):143-56
11325598
Science. 2003 May 30;300(5624):1394-9
12730500
J Virol. 2006 May;80(9):4501-9
16611910
Nat Immunol. 2006 Feb;7(2):131-7
16424890
J Virol. 2005 Dec;79(24):15511-24
16306622
JAMA. 2003 Dec 24;290(24):3222-8
14693875
Annu Rev Biochem. 1998;67:227-64
9759489
J Virol. 2007 Jan;81(1):416-22
17035307
N Engl J Med. 2003 May 15;348(20):1953-66
12690092
J Mol Biol. 2003 Aug 29;331(5):991-1004
12927536
Mol Cells. 2006 Feb 28;21(1):21-8
16511343
FEMS Immunol Med Microbiol. 2006 Mar;46(2):236-43
16487305
Microbes Infect. 2002 May;4(6):647-55
12048034
J Biol Chem. 2006 Apr 21;281(16):10669-81
16431923
J Virol. 2005 Jun;79(12):7819-26
15919935
J Virol. 2005 Aug;79(15):9470-9
16014910
Eur J Biochem. 1992 Jun 15;206(3):901-10
1318841
J Virol. 2005 Feb;79(4):2079-86
15681410
J Virol. 2004 Sep;78(18):9977-86
15331731
Virology. 2006 Jan 5;344(1):119-30
16364743
Virology. 1998 Dec 20;252(2):324-30
9878611
Proc Natl Acad Sci U S A. 2003 Oct 28;100(22):12995-3000
14569023
J Virol. 2007 Jan;81(2):548-57
17108024
Nature. 2003 May 15;423(6937):240
12748632
Biochimie. 2007 Jun-Jul;89(6-7):836-42
17336442
J Virol. 2003 Dec;77(24):13257-66
14645582
J Immunol Methods. 1995 Jul 17;184(1):39-51
7622868
Virology. 2006 Mar 1;346(1):74-85
16303160
J Virol. 2005 Nov;79(21):13800-5
16227300
Curr Opin Immunol. 2000 Aug;12(4):419-24
10899033
Nat Med. 2004 Mar;10(3):290-3
14981511
J Interferon Cytokine Res. 2004 Jul;24(7):388-90
15296649
J Infect Dis. 2004 Apr 1;189(7):1164-7
15031783
Cell. 1995 Dec 29;83(7):1091-100
8548797
J Virol. 2004 Oct;78(20):11416-21
15452265
Science. 2003 May 30;300(5624):1399-404
12730501
Curr Protoc Cytom. 2001 May;Chapter 7:Unit 7.5
18770732
Swiss Med Wkly. 2003 Nov 22;133(43-44):586-90
14745653
Lancet. 2003 Apr 19;361(9366):1319-25
12711465
Viral Immunol. 2004;17(4):498-515
15671747
J Virol. 2005 Dec;79(24):15199-208
16306591
Mol Cell. 1998 Mar;1(4):507-18
9660935
Annu Rev Microbiol. 2001;55:255-81
11544356
Eur J Biochem. 2002 Dec;269(24):6142-51
12473110