PMID- 19144713
OWN - NLM
STAT- MEDLINE
DCOM- 20090330
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 83
IP  - 7
DP  - 2009 Apr
TI  - 5' cis elements direct nodavirus RNA1 recruitment to mitochondrial sites of 
      replication complex formation.
PG  - 2976-88
LID - 10.1128/JVI.02040-08 [doi]
AB  - Positive-strand RNA viruses replicate their genomes on intracellular membranes, 
      usually in conjunction with virus-induced membrane rearrangements. For the 
      nodavirus flock house virus (FHV), we recently showed that multifunctional FHV 
      replicase protein A induces viral RNA template recruitment to a 
      membrane-associated state, but the site(s) and function of this recruitment were 
      not determined. By tagging viral RNA with green fluorescent protein, we show here 
      in Drosophila cells that protein A recruits FHV RNA specifically to the outer 
      mitochondrial membrane sites of RNA replication complex formation. Using 
      Drosophila cells and yeast cells, which also support FHV replication, we also 
      defined the cis-acting regions that direct replication and template recruitment 
      for FHV genomic RNA1. RNA1 nucleotides 68 to 205 were required for RNA 
      replication and directed efficient protein A-mediated RNA recruitment in both 
      cell types. RNA secondary structure prediction, structure probing, and 
      phylogenetic comparisons in this region identified two stable, conserved 
      stem-loops with nearly identical loop sequences. Further mutational analysis 
      showed that both stem-loops and certain flanking sequences were required for RNA1 
      recruitment, negative-strand synthesis, and subsequent positive-strand 
      amplification in yeast and Drosophila cells. Thus, we have shown that protein A 
      recruits RNA1 templates to mitochondria, as expected for RNA replication, and 
      identified a new RNA1 cis element that is necessary and sufficient for RNA1 
      template recognition and recruitment to these mitochondrial membranes for 
      negative-strand RNA1 synthesis. These results establish RNA recruitment to the 
      sites of replication complex formation as an essential, distinct, and selective 
      early step in nodavirus replication.
FAU - Van Wynsberghe, Priscilla M
AU  - Van Wynsberghe PM
AD  - Institute for Molecular Virology, University of Wisconsin-Madison, Madison, 
      Wisconsin 53706-1596, USA.
FAU - Ahlquist, Paul
AU  - Ahlquist P
LA  - eng
GR  - R01 GM035072/GM/NIGMS NIH HHS/United States
GR  - T32 GM007215/GM/NIGMS NIH HHS/United States
GR  - GM35072/GM/NIGMS NIH HHS/United States
GR  - R37 GM035072/GM/NIGMS NIH HHS/United States
GR  - HHMI/Howard Hughes Medical Institute/United States
GR  - T32 GM07215/GM/NIGMS NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20090114
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Proteins)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Drosophila
MH  - Mitochondria/*virology
MH  - Nodaviridae/*physiology
MH  - Nucleic Acid Conformation
MH  - Phylogeny
MH  - RNA, Viral/*genetics/*metabolism
MH  - Saccharomyces cerevisiae/virology
MH  - Sequence Homology, Nucleic Acid
MH  - Viral Proteins/metabolism
MH  - *Virus Replication
PMC - PMC2655588
EDAT- 2009/01/16 09:00
MHDA- 2009/03/31 09:00
PMCR- 2009/10/01
CRDT- 2009/01/16 09:00
PHST- 2009/01/16 09:00 [entrez]
PHST- 2009/01/16 09:00 [pubmed]
PHST- 2009/03/31 09:00 [medline]
PHST- 2009/10/01 00:00 [pmc-release]
AID - JVI.02040-08 [pii]
AID - 2040-08 [pii]
AID - 10.1128/JVI.02040-08 [doi]
PST - ppublish
SO  - J Virol. 2009 Apr;83(7):2976-88. doi: 10.1128/JVI.02040-08. Epub 2009 Jan 14.