PMID- 14599798
OWN - NLM
STAT- MEDLINE
DCOM- 20031211
LR  - 20200409
IS  - 0042-6822 (Print)
IS  - 1096-0341 (Electronic)
IS  - 0042-6822 (Linking)
VI  - 316
IP  - 1
DP  - 2003 Nov 10
TI  - Peptide domains involved in the localization of the porcine reproductive and 
      respiratory syndrome virus nucleocapsid protein to the nucleolus.
PG  - 135-45
AB  - The nucleocapsid (N) protein of porcine reproductive and respiratory syndrome 
      virus (PRRSV) is the principal component of the viral nucleocapsid and localizes 
      to the nucleolus. Peptide sequence analysis of the N protein of several North 
      American isolates identified two potential nuclear localization signal (NLS) 
      sequences located at amino acids 10-13 and 41-42, which were labeled NLS-1 and 
      NLS-2, respectively. Peptides containing NLS-1 or NLS-2 were sufficient to 
      accumulate enhanced green fluorescent protein (EGFP) in the nucleus. The 
      inactivation of NLS-1 by site-directed mutagenesis or the deletion of the first 
      14 amino acids did not affect N protein localization to the nucleolus. The 
      substitution of key lysine residues with uncharged amino acids in NLS-2 blocked 
      nuclear/nucleolar localization. Site-directed mutagenesis within NLS-2 identified 
      the sequence, KKNKK, as forming the core localization domain within NLS-2. Using 
      an in vitro pull-down assay, the N protein was able to bind importin-alpha, 
      importin-beta nuclear transport proteins. The localization pattern of N-EGFP 
      fusion peptides represented by a series of deletions from the C- and N-terminal 
      ends of the N protein identified a region covering amino acids 41-72, which 
      contained a nucleolar localization signal (NoLS) sequence. The 41-72 N peptide 
      when fused to EGFP mimicked the nucleolar-cytoplasmic distribution of native N. 
      These results identify a single NLS involved in the transport of N from the 
      cytoplasm and into nucleus. An additional peptide sequence, overlapping NLS-2, is 
      involved in the further targeting of N to the nucleolus.
FAU - Rowland, Raymond R R
AU  - Rowland RR
AD  - Department of Diagnostic Medicine and Pathobiology, 1800 Denison Avenue, Kansas 
      State University, Manhattan, KS 66506, USA. browland@vet.ksu.edu
FAU - Schneider, Paula
AU  - Schneider P
FAU - Fang, Ying
AU  - Fang Y
FAU - Wootton, Sarah
AU  - Wootton S
FAU - Yoo, Dongwan
AU  - Yoo D
FAU - Benfield, David A
AU  - Benfield DA
LA  - eng
GR  - R15GM61319-01/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Virology
JT  - Virology
JID - 0110674
RN  - 0 (Luminescent Proteins)
RN  - 0 (Nuclear Localization Signals)
RN  - 0 (Nucleocapsid Proteins)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Active Transport, Cell Nucleus
MH  - Amino Acid Sequence
MH  - Animals
MH  - Cell Line
MH  - Cell Nucleolus/metabolism/*virology
MH  - Gene Expression Regulation, Viral
MH  - Green Fluorescent Proteins
MH  - Luminescent Proteins/genetics/metabolism
MH  - Molecular Sequence Data
MH  - *Nuclear Localization Signals
MH  - Nucleocapsid Proteins/*chemistry/genetics/metabolism
MH  - Porcine respiratory and reproductive syndrome virus/*physiology
MH  - Recombinant Fusion Proteins/genetics/metabolism
PMC - PMC7125632
EDAT- 2003/11/06 05:00
MHDA- 2003/12/12 05:00
PMCR- 2003/10/29
CRDT- 2003/11/06 05:00
PHST- 2003/11/06 05:00 [pubmed]
PHST- 2003/12/12 05:00 [medline]
PHST- 2003/11/06 05:00 [entrez]
PHST- 2003/10/29 00:00 [pmc-release]
AID - S0042682203004823 [pii]
AID - S0042-6822(03)00482-3 [pii]
AID - 10.1016/s0042-6822(03)00482-3 [doi]
PST - ppublish
SO  - Virology. 2003 Nov 10;316(1):135-45. doi: 10.1016/s0042-6822(03)00482-3.