Identification of FBL2 as a geranylgeranylated required for hepatitis C cellular protein virus RNA replication

被引:236
作者
Wang, CF
Gale, M
Keller, BC
Huang, H
Brown, MS
Goldstein, JL [1 ]
Ye, J
机构
[1] Univ Texas, SW Med Ctr, Dept Mol Genet, Dallas, TX 75390 USA
[2] Univ Texas, SW Med Ctr, Dept Microbiol, Dallas, TX 75390 USA
关键词
D O I
10.1016/j.molcel.2005.04.004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
We recently reported that Hepatitis C virus (HCV) RNA replication requires one or more geranylgeranylated host proteins. Using a combination of [H-3]mevalonate labeling, coimmunoprecipitation, and bioinformatic search, we identified a geranylgeranylated host protein required for HCV RNA replication. This protein, FBL2, contains an F box domain and a CAAX motif (CVIL). It forms a stable immunoprecipitable complex with the HCV nonstructural protein 5A (NS5A). The association of FBL2 with NS5A requires the CAAX motif of FBL2, but not the F box. Deletion of the F box created a dominant-negative protein that inhibited replication of HCV RNA when overexpressed in Huh7-K2040 cells; this inhibition was overcome by coexpression of NS5A. siRNA-mediated knockdown of FBL2 mRNA by 70% in Huh7-HP cells reduced HCV RNA by 65%; this reduction was overcome by expression of a cDNA encoding a wobble mutant of FBL2. The current data indicate that geranylgeranylated FBL2 binds to NS5A in a reaction crucial for HCV RNA replication.
引用
收藏
页码:425 / 434
页数:10
相关论文
共 41 条
[1]   Cholesterol and 25-hydroxycholesterol inhibit activation of SREBPs by different mechanisms, both involving SCAP and insigs [J].
Adams, CM ;
Reitz, J ;
De Brabander, JK ;
Feramisco, JD ;
Li, L ;
Brown, MS ;
Goldstein, JL .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2004, 279 (50) :52772-52780
[2]  
[Anonymous], FIELDS VIROLOGY
[3]   STRUCTURE OF 2 FORMS OF THE INTERFERON-INDUCED (2-'-5-') OLIGO-A SYNTHETASE OF HUMAN-CELLS BASED ON CDNAS AND GENE-SEQUENCES [J].
BENECH, P ;
MORY, Y ;
REVEL, M ;
CHEBATH, J .
EMBO JOURNAL, 1985, 4 (09) :2249-2256
[4]  
BERNIER L, 1987, J NEUROSCI, V7, P2703
[5]   An amino-terminal amphipathic α-helix mediates membrane association of the hepatitis C virus nonstructural protein 5A [J].
Brass, V ;
Bieck, E ;
Montserret, R ;
Wölk, B ;
Hellings, JA ;
Blum, HE ;
Penin, F ;
Moradpour, D .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (10) :8130-8139
[6]   The molecular biology of West Nile virus: A new invader of the Western hemisphere [J].
Brinton, MA .
ANNUAL REVIEW OF MICROBIOLOGY, 2002, 56 :371-402
[7]   The SCF ubiquitin ligase: Insights into a molecular machine [J].
Cardozo, T ;
Pagano, M .
NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2004, 5 (09) :739-751
[8]   Synaptotagmin gene content of the sequenced genomes [J].
Craxton, M .
BMC GENOMICS, 2004, 5 (1)
[9]   Expression of hepatitis C virus proteins induces distinct membrane alterations including a candidate viral replication complex [J].
Egger, D ;
Wölk, B ;
Gosert, R ;
Bianchi, L ;
Blum, HE ;
Moradpour, D ;
Bienz, K .
JOURNAL OF VIROLOGY, 2002, 76 (12) :5974-5984
[10]   Replication of hepatitis C virus RNA occurs in a membrane-bound replication complex containing nonstructural viral proteins and RNA [J].
El-Hage, N ;
Luo, GX .
JOURNAL OF GENERAL VIROLOGY, 2003, 84 :2761-2769