PMID- 23761406
OWN - NLM
STAT- MEDLINE
DCOM- 20131106
LR  - 20200226
IS  - 1465-2099 (Electronic)
IS  - 0022-1317 (Linking)
VI  - 94
IP  - Pt 9
DP  - 2013 Sep
TI  - Targeted mutations in a highly conserved motif of the nsp1beta protein impair the 
      interferon antagonizing activity of porcine reproductive and respiratory syndrome 
      virus.
PG  - 1972-1983
LID - 10.1099/vir.0.051748-0 [doi]
AB  - Non-structural protein 1beta (nsp1beta) of porcine reproductive and respiratory 
      syndrome virus (PRRSV) contains a papain-like cysteine protease (PLPbeta) domain and 
      has been identified as the main viral protein antagonizing the host innate immune 
      response. In this study, nsp1beta was determined to suppress the expression of 
      reporter genes as well as to suppress 'self-expression' in transfected cells, and 
      this activity appeared to be associated with its interferon (IFN) antagonist 
      function. To knock down the effect of nsp1beta on IFN activity, a panel of 
      site-specific mutations in nsp1beta was analysed. Double mutations K130A/R134A (type 
      1 PRRSV) or K124A/R128A (type 2 PRRSV) targeting a highly conserved motif of 
      nsp1beta, GKYLQRRLQ (in bold), impaired the ability of nsp1beta to suppress IFN-beta and 
      reporter gene expression, as well as to suppress 'self-expression' in vitro. 
      Subsequently, viable recombinant viruses vSD01-08-K130A/R134A and 
      vSD95-21-K124A/R128A, containing double mutations in the GKYLQRRLQ motif were 
      generated using reverse genetics. In comparison with WT viruses, these nsp1beta 
      mutants showed impaired growth ability in infected cells, but the PLPbeta cleavage 
      function was not directly affected. The expression of selected innate immune 
      genes was determined in vSD95-21-K124A/R128A mutant-infected cells. The results 
      consistently showed that gene expression levels of IFN-alpha, IFN-beta and 
      IFN-stimulated gene 15 were upregulated in cells that were infected with the 
      vSD95-21-K124A/R128A compared with that of WT virus. These data suggest that 
      PRRSV nsp1beta may selectively suppress cellular gene expression, including 
      expression of genes involved in the host innate immune function. Modifying the 
      key residues in the highly conserved GKYLQRRLQ motif could attenuate virus growth 
      and improve the cellular innate immune responses.
FAU - Li, Yanhua
AU  - Li Y
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD 57007, USA.
FAU - Zhu, Longchao
AU  - Zhu L
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD 57007, USA.
FAU - Lawson, Steven R
AU  - Lawson SR
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD 57007, USA.
FAU - Fang, Ying
AU  - Fang Y
AD  - Department of Biology/Microbiology, South Dakota State University, Brookings, SD 
      57007, USA.
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD 57007, USA.
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20130612
PL  - England
TA  - J Gen Virol
JT  - The Journal of general virology
JID - 0077340
RN  - 0 (Mutant Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 9008-11-1 (Interferons)
RN  - EC 3.4.- (Cysteine Proteases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Animals
MH  - Cell Line
MH  - Cysteine Proteases/genetics/*metabolism
MH  - DNA Mutational Analysis
MH  - *Host-Pathogen Interactions
MH  - Humans
MH  - Immune Evasion
MH  - Interferons/*antagonists & inhibitors/metabolism
MH  - Macrophages/virology
MH  - Molecular Sequence Data
MH  - Mutagenesis, Site-Directed
MH  - Mutant Proteins/genetics/metabolism
MH  - Porcine respiratory and reproductive syndrome virus/genetics/growth & 
      development/*immunology
MH  - Reverse Genetics
MH  - Viral Nonstructural Proteins/genetics/*metabolism
EDAT- 2013/06/14 06:00
MHDA- 2013/11/07 06:00
CRDT- 2013/06/14 06:00
PHST- 2013/06/14 06:00 [entrez]
PHST- 2013/06/14 06:00 [pubmed]
PHST- 2013/11/07 06:00 [medline]
AID - 10.1099/vir.0.051748-0 [doi]
PST - ppublish
SO  - J Gen Virol. 2013 Sep;94(Pt 9):1972-1983. doi: 10.1099/vir.0.051748-0. Epub 2013 
      Jun 12.