PMID- 19357161
OWN - NLM
STAT- MEDLINE
DCOM- 20090528
LR  - 20211020
IS  - 1098-5514 (Electronic)
IS  - 0022-538X (Print)
IS  - 0022-538X (Linking)
VI  - 83
IP  - 12
DP  - 2009 Jun
TI  - Identification of a novel determinant for membrane association in hepatitis C 
      virus nonstructural protein 4B.
PG  - 6257-68
LID - 10.1128/JVI.02663-08 [doi]
AB  - Nonstructural protein 4B (NS4B) plays an essential role in the formation of the 
      hepatitis C virus (HCV) replication complex. It is a relatively poorly 
      characterized integral membrane protein predicted to comprise four transmembrane 
      segments in its central portion. Here, we describe a novel determinant for 
      membrane association represented by amino acids (aa) 40 to 69 in the N-terminal 
      portion of NS4B. This segment was sufficient to target and tightly anchor the 
      green fluorescent protein to cellular membranes, as assessed by fluorescence 
      microscopy as well as membrane extraction and flotation analyses. Circular 
      dichroism and nuclear magnetic resonance structural analyses showed that this 
      segment comprises an amphipathic alpha-helix extending from aa 42 to 66. 
      Attenuated total reflection infrared spectroscopy and glycosylation acceptor site 
      tagging revealed that this amphipathic alpha-helix has the potential to traverse 
      the phospholipid bilayer as a transmembrane segment, likely upon oligomerization. 
      Alanine substitution of the fully conserved aromatic residues on the hydrophobic 
      helix side abrogated membrane association of the segment comprising aa 40 to 69 
      and disrupted the formation of a functional replication complex. These results 
      provide the first atomic resolution structure of an essential membrane-associated 
      determinant of HCV NS4B.
FAU - Gouttenoire, Jerome
AU  - Gouttenoire J
AD  - Division of Gastroenterology and Hepatology, Centre Hospitalier Universitaire 
      Vaudois, University of Lausanne, Lausanne, Switzerland.
FAU - Castet, Valerie
AU  - Castet V
FAU - Montserret, Roland
AU  - Montserret R
FAU - Arora, Naveen
AU  - Arora N
FAU - Raussens, Vincent
AU  - Raussens V
FAU - Ruysschaert, Jean-Marie
AU  - Ruysschaert JM
FAU - Diesis, Eric
AU  - Diesis E
FAU - Blum, Hubert E
AU  - Blum HE
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 - 20090408
PL  - United States
TA  - J Virol
JT  - Journal of virology
JID - 0113724
RN  - 0 (NS4B protein, flavivirus)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 0 (Viral Nonstructural Proteins)
RN  - 147336-22-9 (Green Fluorescent Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Amino Acid Substitution
MH  - Cell Line, Tumor
MH  - Cell Membrane/*virology
MH  - Circular Dichroism
MH  - Green Fluorescent Proteins/chemistry
MH  - Hepacivirus/*genetics
MH  - Humans
MH  - Magnetic Resonance Spectroscopy
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Protein Structure, Secondary
MH  - Protein Structure, Tertiary
MH  - Recombinant Fusion Proteins/chemistry
MH  - Sequence Alignment
MH  - Spectroscopy, Fourier Transform Infrared
MH  - Viral Nonstructural Proteins/*chemistry
PMC - PMC2687391
EDAT- 2009/04/10 09:00
MHDA- 2009/05/29 09:00
PMCR- 2009/12/01
CRDT- 2009/04/10 09:00
PHST- 2009/04/10 09:00 [entrez]
PHST- 2009/04/10 09:00 [pubmed]
PHST- 2009/05/29 09:00 [medline]
PHST- 2009/12/01 00:00 [pmc-release]
AID - JVI.02663-08 [pii]
AID - 2663-08 [pii]
AID - 10.1128/JVI.02663-08 [doi]
PST - ppublish
SO  - J Virol. 2009 Jun;83(12):6257-68. doi: 10.1128/JVI.02663-08. Epub 2009 Apr 8.