Proteomic profiling of lipid droplet proteins in hepatoma cell lines expressing hepatitis C virus core protein

被引:140
作者
Sato, Shigeko
Fukasawa, Masayoshi [1 ]
Yamakawa, Yoshio
Natsume, Tohru
Suzuki, Tetsuro
Shoji, Ikuo
Aizaki, Hideki
Miyamura, Tatsuo
Nishijima, Masahiro
机构
[1] Natl Inst Infect Dis, Dept Biochem & Cell Biol, Tokyo 1628640, Japan
[2] Natl Inst Infect Dis, Dept Virol 2, Tokyo 1628640, Japan
[3] AIST, Biol Informat Res Ctr, Tokyo 1350064, Japan
关键词
ADRP; DEAD box protein; hepatitis C virus; lipid droplet; TIP47;
D O I
10.1093/jb/mvj104
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
Hepatitis C virus (HCV) core protein has been suggested to play crucial roles in the pathogeneses of liver steatosis and hepatocellular carcinomas due to HCV infection. Intracellular HCV core protein is localized mainly in lipid droplets, in which the core protein should exert its significant biological/pathological functions. In this study, we performed comparative proteomic analysis of lipid droplet proteins in core-expressing and non-expressing hepatoma cell lines. We identified 38 proteins in the lipid droplet fraction of core-expressing (Hep39) cells and 30 proteins in that of non-expressing (Hepswx) cells by 1-D-SDS-PAGE/MALDI-TOF mass spectrometry (MS) or direct nano-flow liquid chromatography-MS/MS. Interestingly, the lipid droplet fraction of Hep39 cells had an apparently lower content of adipose differentiation-related protein and a much higher content of TIP47 than that of Hepswx cells, suggesting the participation of the core protein in lipid droplet biogenesis in HCV-infected cells. Another distinct feature is that proteins involved in RNA metabolism, particularly DEAD box protein 1 and DEAD box protein 3, were detected in the lipid droplet fraction of Hep39 cells. These results suggest that lipid droplets containing HCV core protein may participate in the RNA metabolism of the host and/or HCV, affecting the pathopoiesis and/or virus replication/production in HCV-infected cells.
引用
收藏
页码:921 / 930
页数:10
相关论文
共 80 条
[11]   Direct interaction of hepatitis C virus core protein with the cellular lymphotoxin-beta receptor modulates the signal pathway of the lymphotoxin-beta receptor [J].
Chen, CM ;
You, LR ;
Hwang, LH ;
Lee, YHW .
JOURNAL OF VIROLOGY, 1997, 71 (12) :9417-9426
[12]   An RNA helicase, DDX1, interacting with poly(A) RNA and heterogeneous nuclear ribonucleoprotein K [J].
Chen, HC ;
Lin, WC ;
Tsay, YG ;
Lee, SC ;
Chang, CJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2002, 277 (43) :40403-40409
[13]   ISOLATION OF A CDNA CLONE DERIVED FROM A BLOOD-BORNE NON-A, NON-B VIRAL-HEPATITIS GENOME [J].
CHOO, QL ;
KUO, G ;
WEINER, AJ ;
OVERBY, LR ;
BRADLEY, DW ;
HOUGHTON, M .
SCIENCE, 1989, 244 (4902) :359-362
[14]   TIP47:: A cargo selection device for mannose 6-phosphate receptor trafficking [J].
Díaz, E ;
Pfeffer, SR .
CELL, 1998, 93 (03) :433-443
[15]  
Dvorak AM, 2003, HISTOL HISTOPATHOL, V18, P943, DOI 10.14670/HH-18.943
[16]   Identification of major proteins in the lipid droplet-enriched fraction isolated from the human hepatocyte cell line HuH7 [J].
Fujimoto, Y ;
Itabe, H ;
Sakai, J ;
Makita, M ;
Noda, J ;
Mori, M ;
Higashi, Y ;
Kojima, S ;
Takano, T .
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH, 2004, 1644 (01) :47-59
[17]   Overexpression of a DEAD box protein (DDX1) in neuroblastoma and retinoblastoma cell lines [J].
Godbout, R ;
Packer, M ;
Bie, WJ .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (33) :21161-21168
[18]   EXPRESSION AND IDENTIFICATION OF HEPATITIS C VIRUS POLYPROTEIN CLEAVAGE PRODUCTS [J].
GRAKOUI, A ;
WYCHOWSKI, C ;
LIN, C ;
FEINSTONE, SM ;
RICE, CM .
JOURNAL OF VIROLOGY, 1993, 67 (03) :1385-1395
[19]   ISOLATION OF CDNAS FOR PERILIPIN-A AND PERILIPIN-B - SEQUENCE AND EXPRESSION OF LIPID DROPLET-ASSOCIATED PROTEINS OF ADIPOCYTES [J].
GREENBERG, AS ;
EGAN, JJ ;
WEK, SA ;
MOOS, MC ;
LONDOS, C ;
KIMMEL, AR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (24) :12035-12039
[20]   An3 protein encoded by a localized maternal mRNA in Xenopus laevis is an ATPase with substrate-specific RNA helicase activity [J].
Gururajan, R ;
Weeks, DL .
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1997, 1350 (02) :169-182