PMID- 17301137
OWN - NLM
STAT- MEDLINE
DCOM- 20070926
LR  - 20201209
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 81
IP  - 9
DP  - 2007 May
TI  - Nodavirus RNA replication protein a induces membrane association of genomic RNA.
PG  - 4633-44
AB  - Positive-strand RNA virus genome replication occurs in membrane-associated RNA 
      replication complexes, whose assembly remains poorly understood. Here we show 
      that prior to RNA replication, the multifunctional, transmembrane RNA replication 
      protein A of the nodavirus flock house virus (FHV) recruits FHV genomic RNA1 to a 
      membrane-associated state in both Drosophila melanogaster and Saccharomyces 
      cerevisiae cells. Protein A has mitochondrial membrane-targeting, 
      self-interaction, RNA-dependent RNA polymerase (RdRp), and RNA capping domains. 
      In the absence of RdRp activity due to an active site mutation (A(D692E)), 
      protein A stimulated RNA1 accumulation by increasing RNA1 stability. Protein 
      A(D692E) stimulated RNA1 accumulation in wild-type cells and in xrn1(-) yeast 
      defective in decapped RNA decay, showing that increased RNA1 stability was not 
      due to protein A-mediated RNA1 recapping. Increased RNA1 stability was closely 
      linked with protein A-induced membrane association of the stabilized RNA and was 
      highly selective for RNA1. Substantial N- and C-proximal regions of protein A 
      were dispensable for these activities. However, increased RNA1 accumulation was 
      eliminated by deleting protein A amino acids (aa) 1 to 370 but was restored 
      completely by adding back the transmembrane domain (aa 1 to 35) and partially by 
      adding back peripheral membrane association sequences in aa 36 to 370. Moreover, 
      although RNA polymerase activity was not required, even small deletions in or 
      around the RdRp domain abolished increased RNA1 accumulation. These and other 
      results show that prior to negative-strand RNA synthesis, multiple domains of 
      mitochondrially targeted protein A cooperate to selectively recruit FHV genomic 
      RNA to membranes where RNA replication complexes form.
FAU - Van Wynsberghe, Priscilla M
AU  - Van Wynsberghe PM
AD  - Institute for Molecular Virology, University of Wisconsin-Madison, 1525 Linden 
      Drive, Madison, WI 53706-1596, USA.
FAU - Chen, Hau-Ren
AU  - Chen HR
FAU - Ahlquist, Paul
AU  - Ahlquist P
LA  - eng
GR  - R01 GM035072/GM/NIGMS NIH HHS/United States
GR  - R37 GM035072/GM/NIGMS NIH HHS/United States
GR  - T32 GM007215/GM/NIGMS NIH HHS/United States
GR  - T32 GM 07215/GM/NIGMS NIH HHS/United States
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20070214
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (RNA, Viral)
RN  - 0 (Replication Protein A)
RN  - EC 2.7.7.48 (RNA-Dependent RNA Polymerase)
SB  - IM
MH  - Animals
MH  - Blotting, Northern
MH  - Blotting, Western
MH  - Cells, Cultured
MH  - Drosophila melanogaster
MH  - Genomic Instability/genetics
MH  - Mutation/genetics
MH  - Nodaviridae/*genetics
MH  - Protein Structure, Tertiary
MH  - RNA, Viral/genetics/*metabolism
MH  - RNA-Dependent RNA Polymerase/genetics
MH  - Replication Protein A/genetics/*metabolism
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Saccharomyces cerevisiae
MH  - Virus Replication/genetics/*physiology
PMC - PMC1900146
EDAT- 2007/02/16 09:00
MHDA- 2007/09/27 09:00
PMCR- 2007/09/01
CRDT- 2007/02/16 09:00
PHST- 2007/02/16 09:00 [pubmed]
PHST- 2007/09/27 09:00 [medline]
PHST- 2007/02/16 09:00 [entrez]
PHST- 2007/09/01 00:00 [pmc-release]
AID - JVI.02267-06 [pii]
AID - 2267-06 [pii]
AID - 10.1128/JVI.02267-06 [doi]
PST - ppublish
SO  - J Virol. 2007 May;81(9):4633-44. doi: 10.1128/JVI.02267-06. Epub 2007 Feb 14.