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Journal of Biochemistry 2006 139(5):921-930; doi:10.1093/jb/mvj104
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© 2006 The Japanese Biochemical Society.

Regular Paper

Proteomic Profiling of Lipid Droplet Proteins in Hepatoma Cell Lines Expressing Hepatitis C Virus Core Protein

Shigeko Sato1, Masayoshi Fukasawa1,*, Yoshio Yamakawa1, Tohru Natsume2, Tetsuro Suzuki3, Ikuo Shoji3, Hideki Aizaki3, Tatsuo Miyamura3 and Masahiro Nishijima1,{dagger}

1 Department of Biochemistry and Cell Biology and 3 Department of Virology II, National Institute of Infectious Diseases, Tokyo 162-8640; and 2 National Institute of Advanced Industrial Science and Technology (AIST), Biological Information Research Center, Tokyo 135-0064

* To whom correspondence should be addressed. Tel: +81-3-5285-1111, Fax: +81-3-5285-1157, E-mail: fuka{at}nih.go.jp

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 nanoflow 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.

{dagger} Present address: Department of Clinical Pharmacy, Faculty of Pharmaceutical Sciences, Doshisha Women's College of Liberal Arts, Kyoto 610-0395.


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