Identification of a novel determinant for membrane association in hepatitis C virus nonstructural protein 4B.

Details

Serval ID
serval:BIB_066B404C2895
Type
Article: article from journal or magazin.
Collection
Publications
Title
Identification of a novel determinant for membrane association in hepatitis C virus nonstructural protein 4B.
Journal
Journal of Virology
Author(s)
Gouttenoire J., Castet V., Montserret R., Arora N., Raussens V., Ruysschaert J.M., Diesis E., Blum H.E., Penin F., Moradpour D.
ISSN
1098-5514[electronic]
Publication state
Published
Issued date
2009
Volume
83
Number
12
Pages
6257-6268
Language
english
Abstract
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.
Keywords
Amino Acid Sequence, Amino Acid Substitution, Cell Line, Tumor, Cell Membrane/virology, Circular Dichroism, Green Fluorescent Proteins/chemistry, Hepacivirus/genetics, Humans, Magnetic Resonance Spectroscopy, Models, Molecular, Molecular Sequence Data, Protein Structure, Secondary, Protein Structure, Tertiary, Recombinant Fusion Proteins/chemistry, Sequence Alignment, Spectroscopy, Fourier Transform Infrared, Viral Nonstructural Proteins/chemistry
Pubmed
Web of science
Open Access
Yes
Create date
27/06/2009 14:54
Last modification date
20/08/2019 12:28
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