PMID- 24089566
OWN - NLM
STAT- MEDLINE
DCOM- 20140205
LR  - 20211021
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 87
IP  - 24
DP  - 2013 Dec
TI  - Highly divergent strains of porcine reproductive and respiratory syndrome virus 
      incorporate multiple isoforms of nonstructural protein 2 into virions.
PG  - 13456-65
LID - 10.1128/JVI.02435-13 [doi]
AB  - Viral structural proteins form the critical intermediary between viral infection 
      cycles within and between hosts, function to initiate entry, participate in 
      immediate early viral replication steps, and are major targets for the host 
      adaptive immune response. We report the identification of nonstructural protein 2 
      (nsp2) as a novel structural component of the porcine reproductive and 
      respiratory syndrome virus (PRRSV) particle. A set of custom alpha-nsp2 antibodies 
      targeting conserved epitopes within four distinct regions of nsp2 (the PLP2 
      protease domain [OTU], the hypervariable domain [HV], the putative transmembrane 
      domain [TM], and the C-terminal region [C]) were obtained commercially and 
      validated in PRRSV-infected cells. Highly purified cell-free virions of several 
      PRRSV strains were isolated through multiple rounds of differential density 
      gradient centrifugation and analyzed by immunoelectron microscopy (IEM) and 
      Western blot assays using the alpha-nsp2 antibodies. Purified viral preparations were 
      found to contain pleomorphic, predominantly spherical virions of uniform size 
      (57.9 nm +/- 8.1 nm diameter; n = 50), consistent with the expected size of PRRSV 
      particles. Analysis by IEM indicated the presence of nsp2 associated with the 
      viral particle of diverse strains of PRRSV. Western blot analysis confirmed the 
      presence of nsp2 in purified viral samples and revealed that multiple nsp2 
      isoforms were associated with the virion. Finally, a recombinant PRRSV genome 
      containing a myc-tagged nsp2 was used to generate purified virus, and these 
      particles were also shown to harbor myc-tagged nsp2 isoforms. Together, these 
      data identify nsp2 as a virion-associated structural PRRSV protein and reveal 
      that nsp2 exists in or on viral particles as multiple isoforms.
FAU - Kappes, Matthew A
AU  - Kappes MA
AD  - Virus and Prion Research Unit, USDA-ARS-National Animal Disease Center, Ames, 
      Iowa, USA.
FAU - Miller, Cathy L
AU  - Miller CL
FAU - Faaberg, Kay S
AU  - Faaberg KS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20131002
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (Protein Isoforms)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - *Evolution, Molecular
MH  - Molecular Sequence Data
MH  - Porcine Reproductive and Respiratory Syndrome/*virology
MH  - Porcine respiratory and reproductive syndrome 
      virus/chemistry/classification/genetics/*metabolism
MH  - Protein Isoforms/chemistry/genetics/metabolism
MH  - Sequence Alignment
MH  - Swine
MH  - Viral Nonstructural Proteins/chemistry/genetics/*metabolism
MH  - Virion/chemistry/classification/genetics/*metabolism
PMC - PMC3838290
EDAT- 2013/10/04 06:00
MHDA- 2014/02/06 06:00
PMCR- 2014/06/01
CRDT- 2013/10/04 06:00
PHST- 2013/10/04 06:00 [entrez]
PHST- 2013/10/04 06:00 [pubmed]
PHST- 2014/02/06 06:00 [medline]
PHST- 2014/06/01 00:00 [pmc-release]
AID - JVI.02435-13 [pii]
AID - 02435-13 [pii]
AID - 10.1128/JVI.02435-13 [doi]
PST - ppublish
SO  - J Virol. 2013 Dec;87(24):13456-65. doi: 10.1128/JVI.02435-13. Epub 2013 Oct 2.