PMID- 25768891
OWN - NLM
STAT- MEDLINE
DCOM- 20150803
LR  - 20150518
IS  - 1096-0341 (Electronic)
IS  - 0042-6822 (Linking)
VI  - 481
DP  - 2015 Jul
TI  - Porcine reproductive and respiratory syndrome virus nonstructural protein 2 
      (nsp2) topology and selective isoform integration in artificial membranes.
PG  - 51-62
LID - S0042-6822(15)00040-9 [pii]
LID - 10.1016/j.virol.2015.01.028 [doi]
AB  - The membrane insertion and topology of nonstructural protein 2 (nsp2) of porcine 
      reproductive and respiratory syndrome virus (PRRSV) strain VR-2332 was assessed 
      using a cell free translation system in the presence or absence of artificial 
      membranes. Expression of PRRSV nsp2 in the absence of all other viral factors 
      resulted in the genesis of both full-length nsp2 as well as a select number of 
      C-terminal nsp2 isoforms. Addition of membranes to the translation stabilized the 
      translation reaction, resulting in predominantly full-length nsp2 as assessed by 
      immunoprecipitation. Analysis further showed full-length nsp2 strongly associates 
      with membranes, along with two additional large nsp2 isoforms. Membrane 
      integration of full-length nsp2 was confirmed through high-speed density 
      fractionation, protection from protease digestion, and immunoprecipitation. The 
      results demonstrated that nsp2 integrated into the membranes with an unexpected 
      topology, where the amino (N)-terminal (cytoplasmic) and C-terminal (luminal) 
      domains were orientated on opposite sides of the membrane surface.
CI  - Published by Elsevier Inc.
FAU - Kappes, Matthew A
AU  - Kappes MA
AD  - Virus and Prion Research Unit, USDA-ARS-National Animal Disease Center, Ames, IA, 
      USA; Department of Veterinary Microbiology and Preventive Medicine, College of 
      Veterinary Medicine, Iowa State University, Ames, IA, USA.
FAU - Miller, Cathy L
AU  - Miller CL
AD  - Department of Veterinary Microbiology and Preventive Medicine, College of 
      Veterinary Medicine, Iowa State University, Ames, IA, USA.
FAU - Faaberg, Kay S
AU  - Faaberg KS
AD  - Virus and Prion Research Unit, USDA-ARS-National Animal Disease Center, Ames, IA, 
      USA. Electronic address: kay.faaberg@ars.usda.gov.
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 - 20150311
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
RN  - 0 (Protein Isoforms)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Cell Membrane/*virology
MH  - Porcine Reproductive and Respiratory Syndrome/*virology
MH  - Porcine respiratory and reproductive syndrome 
      virus/chemistry/genetics/*metabolism
MH  - Protein Isoforms/chemistry/genetics/metabolism
MH  - Protein Structure, Tertiary
MH  - Swine
MH  - Viral Nonstructural Proteins/*chemistry/genetics/*metabolism
OTO - NOTNLM
OT  - Arterivirus
OT  - Membrane topology
OT  - Nonstructural protein 2
OT  - Porcine reproductive and respiratory syndrome virus
OT  - Protein isomers
OT  - Structural protein
EDAT- 2015/03/15 06:00
MHDA- 2015/08/04 06:00
CRDT- 2015/03/14 06:00
PHST- 2014/12/06 00:00 [received]
PHST- 2015/01/15 00:00 [revised]
PHST- 2015/01/30 00:00 [accepted]
PHST- 2015/03/14 06:00 [entrez]
PHST- 2015/03/15 06:00 [pubmed]
PHST- 2015/08/04 06:00 [medline]
AID - S0042-6822(15)00040-9 [pii]
AID - 10.1016/j.virol.2015.01.028 [doi]
PST - ppublish
SO  - Virology. 2015 Jul;481:51-62. doi: 10.1016/j.virol.2015.01.028. Epub 2015 Mar 11.