PMID- 20926561
OWN - NLM
STAT- MEDLINE
DCOM- 20110106
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 84
IP  - 24
DP  - 2010 Dec
TI  - Amphipathic alpha-helix AH2 is a major determinant for the oligomerization of 
      hepatitis C virus nonstructural protein 4B.
PG  - 12529-37
LID - 10.1128/JVI.01798-10 [doi]
AB  - Nonstructural protein 4B (NS4B) is a key organizer of hepatitis C virus (HCV) 
      replication complex formation. It induces a specific membrane rearrangement, 
      designated membranous web, that serves as a scaffold for the HCV replication 
      complex. However, the mechanisms underlying membranous web formation are poorly 
      understood. Based on fluorescence resonance energy transfer (FRET) and 
      confirmatory coimmunoprecipitation analyses, we provide evidence for an 
      oligomerization of NS4B in the membrane environment of intact cells. Several 
      conserved determinants were found to be involved in NS4B oligomerization, through 
      homotypic and heterotypic interactions. N-terminal amphipathic alpha-helix AH2, 
      comprising amino acids 42 to 66, was identified as a major determinant for NS4B 
      oligomerization. Mutations that affected the oligomerization of NS4B disrupted 
      membranous web formation and HCV RNA replication, implying that oligomerization 
      of NS4B is required for the creation of a functional replication complex. These 
      findings enhance our understanding of the functional architecture of the HCV 
      replication complex and may provide new angles for therapeutic intervention. At 
      the same time, they expand the list of positive-strand RNA virus replicase 
      components acting as oligomers.
FAU - Gouttenoire, Jerome
AU  - Gouttenoire J
AD  - Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire 
      Vaudois, BU44/07/2421, Rue du Bugnon 44, CH-1011 Lausanne, Switzerland.
FAU - Roingeard, Philippe
AU  - Roingeard P
FAU - Penin, Francois
AU  - Penin F
FAU - Moradpour, Darius
AU  - Moradpour D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20101006
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (RNA, Viral)
RN  - 0 (Viral Nonstructural Proteins)
SB  - IM
MH  - Bone Neoplasms/genetics/metabolism/virology
MH  - Carcinoma, Hepatocellular/genetics/metabolism/virology
MH  - Cell Membrane/virology
MH  - Fluorescence Resonance Energy Transfer
MH  - Hepacivirus/genetics/*metabolism
MH  - Hepatitis C/genetics/*metabolism/virology
MH  - Humans
MH  - Immunoprecipitation
MH  - Liver Neoplasms/genetics/metabolism/virology
MH  - Osteosarcoma/genetics/metabolism/virology
MH  - Plasmids
MH  - *Protein Multimerization
MH  - Protein Structure, Secondary
MH  - RNA, Viral/genetics
MH  - Tumor Cells, Cultured
MH  - Viral Nonstructural Proteins/*chemistry/genetics/*metabolism
MH  - Virus Replication
PMC - PMC3004355
EDAT- 2010/10/12 06:00
MHDA- 2011/01/07 06:00
PMCR- 2011/06/01
CRDT- 2010/10/08 06:00
PHST- 2010/10/08 06:00 [entrez]
PHST- 2010/10/12 06:00 [pubmed]
PHST- 2011/01/07 06:00 [medline]
PHST- 2011/06/01 00:00 [pmc-release]
AID - JVI.01798-10 [pii]
AID - 1798-10 [pii]
AID - 10.1128/JVI.01798-10 [doi]
PST - ppublish
SO  - J Virol. 2010 Dec;84(24):12529-37. doi: 10.1128/JVI.01798-10. Epub 2010 Oct 6.