PMID- 22301142
OWN - NLM
STAT- MEDLINE
DCOM- 20120618
LR  - 20211021
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 86
IP  - 8
DP  - 2012 Apr
TI  - Disulfide linkages mediating nucleocapsid protein dimerization are not required 
      for porcine arterivirus infectivity.
PG  - 4670-81
LID - 10.1128/JVI.06709-11 [doi]
AB  - The nucleocapsid (N) proteins of the North American (type II) and European (type 
      I) genotypes of porcine reproductive and respiratory syndrome virus (PRRSV) share 
      only approximately 60% genetic identity, and the functionality of N in both 
      genotypes, especially its role in virion assembly, is still poorly understood. In 
      this study, we demonstrated that the ORF7 3' untranslated region or ORF7 of type 
      I is functional in the type II PRRSV background. Based on these results, we 
      postulated that the cysteine at position 90 (Cys90) of the type II N protein, 
      which corresponds to an alanine in the type I protein, is nonessential for virus 
      infectivity. The replacement of Cys90 with alanine confirmed this hypothesis. We 
      then hypothesized that all of the cysteines in the N protein could be replaced by 
      alanines. Mutational analysis revealed that, in contradiction to previously 
      reported findings, the replacement of all of the cysteines, either singly or in 
      combination, did not impair the growth of either type II or type I PRRSV. 
      Treatment with the alkylating agent N-ethylmaleimide inhibited cysteine-mediated 
      N dimerization in living cells but not in released virions. Additionally, 
      bimolecular fluorescence complementation assays revealed noncovalent interactions 
      in living cells among the N and C termini and between the N-terminal and 
      C-terminal regions of the N proteins of both genotypes of PRRSV. These results 
      demonstrate that the disulfide linkages mediating the N dimerization are not 
      required for PRRSV viability and help to promote our understanding of the 
      mechanism underlying arterivirus particle assembly.
FAU - Zhang, Rong
AU  - Zhang R
AD  - Department of Swine Infectious Diseases, Shanghai Veterinary Research Institute, 
      The Chinese Academy of Agricultural Sciences, Shanghai 200241, China.
FAU - Chen, Chunyan
AU  - Chen C
FAU - Sun, Zhi
AU  - Sun Z
FAU - Tan, Feifei
AU  - Tan F
FAU - Zhuang, Jinshan
AU  - Zhuang J
FAU - Tian, Debin
AU  - Tian D
FAU - Tong, Guangzhi
AU  - Tong G
FAU - Yuan, Shishan
AU  - Yuan S
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20120201
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (Nucleocapsid Proteins)
RN  - K848JZ4886 (Cysteine)
RN  - O3C74ACM9V (Ethylmaleimide)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cell Line
MH  - Cysteine/chemistry/metabolism
MH  - Ethylmaleimide/pharmacology
MH  - Gene Order
MH  - Molecular Sequence Data
MH  - Mutation
MH  - Nucleocapsid Proteins/*chemistry/genetics/*metabolism
MH  - Porcine respiratory and reproductive syndrome 
      virus/*chemistry/genetics/*metabolism
MH  - Protein Binding
MH  - Protein Multimerization/drug effects
MH  - Sequence Alignment
MH  - Virion/drug effects/metabolism
PMC - PMC3318635
EDAT- 2012/02/04 06:00
MHDA- 2012/06/19 06:00
PMCR- 2012/10/01
CRDT- 2012/02/04 06:00
PHST- 2012/02/04 06:00 [entrez]
PHST- 2012/02/04 06:00 [pubmed]
PHST- 2012/06/19 06:00 [medline]
PHST- 2012/10/01 00:00 [pmc-release]
AID - JVI.06709-11 [pii]
AID - 06709-11 [pii]
AID - 10.1128/JVI.06709-11 [doi]
PST - ppublish
SO  - J Virol. 2012 Apr;86(8):4670-81. doi: 10.1128/JVI.06709-11. Epub 2012 Feb 1.