PMID- 11689648
OWN - NLM
STAT- MEDLINE
DCOM- 20011207
LR  - 20181113
IS  - 0022-538X (Print)
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Linking)
VI  - 75
IP  - 23
DP  - 2001 Dec
TI  - Flock house virus RNA replicates on outer mitochondrial membranes in Drosophila 
      cells.
PG  - 11664-76
AB  - The identification and characterization of host cell membranes essential for 
      positive-strand RNA virus replication should provide insight into the mechanisms 
      of viral replication and potentially identify novel targets for broadly effective 
      antiviral agents. The alphanodavirus flock house virus (FHV) is a positive-strand 
      RNA virus with one of the smallest known genomes among animal RNA viruses, and it 
      can replicate in insect, plant, mammalian, and yeast cells. To investigate the 
      localization of FHV RNA replication, we generated polyclonal antisera against 
      protein A, the FHV RNA-dependent RNA polymerase, which is the sole viral protein 
      required for FHV RNA replication. We detected protein A within 4 h after 
      infection of Drosophila DL-1 cells and, by differential and isopycnic gradient 
      centrifugation, found that protein A was tightly membrane associated, similar to 
      integral membrane replicase proteins from other positive-strand RNA viruses. 
      Confocal immunofluorescence microscopy and virus-specific, actinomycin 
      D-resistant bromo-UTP incorporation identified mitochondria as the intracellular 
      site of protein A localization and viral RNA synthesis. Selective membrane 
      permeabilization and immunoelectron microscopy further localized protein A to 
      outer mitochondrial membranes. Electron microscopy revealed 40- to 60-nm 
      membrane-bound spherical structures in the mitochondrial intermembrane space of 
      FHV-infected cells, similar in ultrastructural appearance to tombusvirus- and 
      togavirus-induced membrane structures. We concluded that FHV RNA replication 
      occurs on outer mitochondrial membranes and shares fundamental biochemical and 
      ultrastructural features with RNA replication of positive-strand RNA viruses from 
      other families.
FAU - Miller, D J
AU  - Miller DJ
AD  - Department of Medicine, University of Wisconsin-Madison, 53706, USA.
FAU - Schwartz, M D
AU  - Schwartz MD
FAU - Ahlquist, P
AU  - Ahlquist P
LA  - eng
GR  - R01 GM035072/GM/NIGMS NIH HHS/United States
GR  - R37 GM035072/GM/NIGMS NIH HHS/United States
GR  - GM35072/GM/NIGMS NIH HHS/United States
GR  - K08 AI0770-01/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, U.S. Gov't, P.H.S.
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  - Blotting, Northern
MH  - Cell Line
MH  - Drosophila melanogaster/cytology/*virology
MH  - Intracellular Membranes/ultrastructure/virology
MH  - Microscopy, Confocal
MH  - Microscopy, Electron/methods
MH  - Microscopy, Fluorescence
MH  - Mitochondria/ultrastructure/virology
MH  - Nodaviridae/*physiology
MH  - RNA, Viral/*biosynthesis
MH  - Subcellular Fractions/metabolism
MH  - Viral Proteins/metabolism
PMC - PMC114753
EDAT- 2001/11/02 10:00
MHDA- 2002/01/05 10:01
PMCR- 2002/06/01
CRDT- 2001/11/02 10:00
PHST- 2001/11/02 10:00 [pubmed]
PHST- 2002/01/05 10:01 [medline]
PHST- 2001/11/02 10:00 [entrez]
PHST- 2002/06/01 00:00 [pmc-release]
AID - 1104 [pii]
AID - 10.1128/JVI.75.23.11664-11676.2001 [doi]
PST - ppublish
SO  - J Virol. 2001 Dec;75(23):11664-76. doi: 10.1128/JVI.75.23.11664-11676.2001.