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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Systematic Identification of Hepatocellular Proteins Interacting with NS5A of the Hepatitis C Virus

  • Ahn, Ji-Won (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Chung, Kyung-Sook (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Dong-Uk (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Won, Mi-Sun (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Li-La (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Kyung-Shin (College of Oriental Medicine, Taejon University) ;
  • Nam, Mi-Young (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Choi, Shin-Jung (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Kim, Hyoung-Chin (Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Yoon, Mi-Chung (Department of Biology, Mokwon University) ;
  • Chae, Suhn-Kee (Department of Biochemistry, Paichai University) ;
  • Hoe, Kwang-Lae (Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • Published : 2004.11.30
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Abstract

The hepatitis C virus is associated with the development of liver cirrhosis and hepatocellular carcinomas. Among the 10 polyproteins produced by the virus, no function has been clearly assigned to the non-structural 5A (NS5A) protein. This study was designed to identify the hepatocellular proteins that interact with NS5A of the HCV. Yeast two-hybrid experiments were performed with a human liver cDNA prey-library, using five different NS5A derivatives as baits, the full-length NS5A (NS5A-F, amino acid (aa) 1~447) and its four different derivatives, denoted as NS5A-A (aa 1~150), -B (aa 1~300), -C (aa 300~447) and D (aa 150~447). NS5A-F, NS5A-B and NS5A-C gave two, two and 10 candidate clones, respectively, including an AHNAK-related protein, the secreted frizzled-related protein 4 (SFRP4), the N-myc downstream regulated gene 1 (NDRG1), the cellular retinoic acid binding protein 1 (CRABP-1), ferritin heavy chain (FTH1), translokin, tumor-associated calcium signal transducer 2 (TACSTD2), phosphatidylinositol 4-kinase (PI4K) and $centaurin{\delta}$ 2 ($CENT{\delta}2$). However, NS5A-A produced no candidates and NS5A-D was not suitable as bait due to transcriptional activity. Based on an in vitro binding assay, CRABP-1, PI4K, $CENT{\delta}2$ and two unknown fusion proteins with maltose binding protein (MBP), were confirmed to interact with the glutathione S-transferase (GST)/NS5A fusion protein. Furthermore, the interactions of CRABP-1, PI4K and $CENT{\delta}2$ were not related to the PXXP motif (class II), as judged by a domain analysis. While their biological relevance is under investigation, the results contribute to a better understanding of the possible role of NS5A in hepatocellular signaling pathways.

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References

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