PMID- 10359774
OWN - NLM
STAT- MEDLINE
DCOM- 19990708
LR  - 20190501
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 96
IP  - 12
DP  - 1999 Jun 8
TI  - Regulation of eukaryotic protein synthesis: selective influenza viral mRNA
      translation is mediated by the cellular RNA-binding protein GRSF-1.
PG  - 6694-9
AB  - To better understand regulation of eukaryotic protein synthesis, we studied
      cellular and viral mRNA translation in influenza virus-infected cells. Influenza 
      virus infection results in a dramatic shut-off of cellular protein synthesis that
      is concomitant with selective viral mRNA translation. Earlier work showed that
      these events are mediated by viral and/or cellular factors binding to the 5'
      untranslated region (5' UTR) of viral mRNAs. To identify trans-acting cellular
      proteins responsible for selective viral protein synthesis, we employed the yeast
      three-hybrid system. Using the 5' UTR of the influenza virus nucleocapsid protein
      (NP) mRNA as bait, we identified the cellular RNA-recognition motif containing
      RNA-binding protein G-rich sequence factor 1 (GRSF-1) as a positive-acting
      translational regulatory factor. The in vivo yeast assay revealed GRSF-1
      specifically bound to the NP 5' UTR but not select NP 5' UTR mutants or cellular 
      RNA 5' UTRs. These data were confirmed by gel shift assays using recombinant
      GRSF-1. Importantly, recombinant GRSF-1 specifically stimulated translation of a 
      NP 5' UTR-driven template in cell-free translation systems. Furthermore,
      translation efficiency of NP 5' UTR-driven templates was reduced markedly in
      GRSF-1-depleted HeLa cell extracts, but restored in GRSF-1-reconstituted
      extracts. GRSF-1 also stimulated translation of an NP 5' UTR-driven template in
      HeLa cell extracts that were depleted of essential factors by addition of RNA
      oligonucleotides representing the viral 5' UTR RNA. Taken together, these data
      document the functional demonstration of a cellular protein binding to influenza 
      virus RNAs and, importantly, suggest that influenza virus may recruit GRSF-1 to
      the 5' UTR to ensure preferential translation of viral mRNAs in infected cells.
FAU - Park, Y W
AU  - Park YW
AD  - Department of Microbiology, School of Medicine, University of Washington,
      Seattle, WA 98195, USA.
FAU - Wilusz, J
AU  - Wilusz J
FAU - Katze, M G
AU  - Katze MG
LA  - eng
GR  - P51 RR000166/RR/NCRR NIH HHS/United States
GR  - RR-00166/RR/NCRR NIH HHS/United States
GR  - GM-56434/GM/NIGMS NIH HHS/United States
GR  - AI-22646/AI/NIAID NIH HHS/United States
GR  - R01 AI022646/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (5' Untranslated Regions)
RN  - 0 (GRSF1 protein, human)
RN  - 0 (Poly(A)-Binding Proteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (RNA, Viral)
RN  - 0 (RNA-Binding Proteins)
SB  - IM
MH  - 5' Untranslated Regions/genetics
MH  - Base Sequence
MH  - Cell-Free System
MH  - HeLa Cells
MH  - Humans
MH  - Influenza, Human/genetics/metabolism
MH  - Molecular Sequence Data
MH  - Orthomyxoviridae/*genetics
MH  - *Poly(A)-Binding Proteins
MH  - *Protein Biosynthesis
MH  - RNA, Messenger/*genetics
MH  - RNA, Viral/genetics
MH  - RNA-Binding Proteins/*genetics/metabolism
PMC - PMC21977
EDAT- 1999/06/09 00:00
MHDA- 1999/06/09 00:01
CRDT- 1999/06/09 00:00
PHST- 1999/06/09 00:00 [pubmed]
PHST- 1999/06/09 00:01 [medline]
PHST- 1999/06/09 00:00 [entrez]
AID - 10.1073/pnas.96.12.6694 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6694-9. doi: 10.1073/pnas.96.12.6694.