PMID- 25557977
OWN - NLM
STAT- MEDLINE
DCOM- 20160216
LR  - 20201209
IS  - 1872-7492 (Electronic)
IS  - 0168-1702 (Linking)
VI  - 202
DP  - 2015 Apr 16
TI  - Proteolytic processing of the porcine reproductive and respiratory syndrome virus 
      replicase.
PG  - 48-59
LID - S0168-1702(14)00541-3 [pii]
LID - 10.1016/j.virusres.2014.12.027 [doi]
AB  - The porcine reproductive and respiratory syndrome virus (PRRSV) replicase 
      polyproteins pp1a and pp1ab are proteolytically processed by four proteases 
      encoded in ORF1a. In this study, a large set of PRRSV replicase cleavage products 
      were identified and pp1a cleavage sites were verified by using a combination of 
      bioinformatics, proteomics, immunoprecipitation, and site-directed mutagenesis. 
      For genotype 1 PRRSV (isolate SD01-08), proteomic analysis identified H180/S181, 
      G385/A386, and G1446/A1447 as the cleavage sites separating nsp1alpha/1beta, nsp1beta/nsp2, 
      and nsp2/nsp3, respectively. Transient expression of nsp2-8, nsp3-8, nsp4-8, 
      nsp5-8 (using the recombinant vaccinia virus/T7 RNA polymerase system) and 
      immunoprecipitation identified the cleavage end products nsp2, nsp3, nsp4, nsp7alpha 
      and nsp7beta, and various processing intermediates. Our studies also revealed the 
      existence of alternative proteolytic processing pathways for the processing of 
      the nsp3-8 region, depending on the presence or absence of nsp2 as a co-factor. 
      The identity of most cleavage products was further corroborated by site-directed 
      mutagenesis of individual cleavage sites in constructs expressing nsp3-8 or 
      nsp4-8. This study constitutes the first in-depth experimental analysis of PRRSV 
      replicase processing and the data are discussed against the background of the 
      processing scheme previously derived for the arterivirus prototype, the distantly 
      related equine arteritis virus (EAV). Despite several differences between the two 
      viruses, of which the functional significance remains to be studied, our study 
      demonstrates the general conservation of the replicase pp1a processing scheme 
      between EAV and PRRSV, and likely also the other members of the arterivirus 
      family.
CI  - Copyright (c) 2014 Elsevier B.V. All rights reserved.
FAU - Li, Yanhua
AU  - Li Y
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD, USA; Department of Diagnostic Medicine and Pathobiology, College 
      of Veterinary Medicine, Kansas State University, Manhattan, KS, USA.
FAU - Tas, Ali
AU  - Tas A
AD  - Department of Medical Microbiology, Center for Infectious Diseases, Leiden 
      University Medical Center, Leiden, The Netherlands.
FAU - Sun, Zhi
AU  - Sun Z
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD, USA.
FAU - Snijder, Eric J
AU  - Snijder EJ
AD  - Department of Medical Microbiology, Center for Infectious Diseases, Leiden 
      University Medical Center, Leiden, The Netherlands. Electronic address: 
      e.j.snijder@lumc.nl.
FAU - Fang, Ying
AU  - Fang Y
AD  - Department of Veterinary and Biomedical Sciences, South Dakota State University, 
      Brookings, SD, USA; Department of Diagnostic Medicine and Pathobiology, College 
      of Veterinary Medicine, Kansas State University, Manhattan, KS, USA. Electronic 
      address: yfang@vet.k-state.edu.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20141231
PL  - Netherlands
TA  - Virus Res
JT  - Virus research
JID - 8410979
RN  - 0 (Viral Proteins)
RN  - EC 2.7.7.48 (RNA-Dependent RNA Polymerase)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Computational Biology
MH  - Immunoprecipitation
MH  - Mutagenesis, Site-Directed
MH  - Porcine respiratory and reproductive syndrome virus/*enzymology/*physiology
MH  - *Protein Processing, Post-Translational
MH  - Proteolysis
MH  - Proteomics
MH  - RNA-Dependent RNA Polymerase/*metabolism
MH  - Viral Proteins/*metabolism
OTO - NOTNLM
OT  - Arterivirus
OT  - Cleavage site
OT  - Nidovirus
OT  - Nonstructural protein
OT  - Protease
OT  - Replication
EDAT- 2015/01/06 06:00
MHDA- 2016/02/18 06:00
CRDT- 2015/01/06 06:00
PHST- 2014/10/14 00:00 [received]
PHST- 2014/12/19 00:00 [revised]
PHST- 2014/12/22 00:00 [accepted]
PHST- 2015/01/06 06:00 [entrez]
PHST- 2015/01/06 06:00 [pubmed]
PHST- 2016/02/18 06:00 [medline]
AID - S0168-1702(14)00541-3 [pii]
AID - 10.1016/j.virusres.2014.12.027 [doi]
PST - ppublish
SO  - Virus Res. 2015 Apr 16;202:48-59. doi: 10.1016/j.virusres.2014.12.027. Epub 2014 
      Dec 31.