Functional interaction of hepatitis C Virus NS5B with Nucleolin GAR domain
| Autor: | Takashi Kusakawa, Tetsuro Shimakami, Katsuji Yoshioka, Seishi Murakami, Shuichi Kaneko |
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| Rok vydání: | 2007 |
| Předmět: |
viruses
Mutant Molecular Sequence Data Biology Viral Nonstructural Proteins Arginine NS5B Biochemistry chemistry.chemical_compound Chlorocebus aethiops Nucleoli Animals Replicon Amino Acid Sequence Amino Acids Molecular Biology Host factor Nucleolin chemistry.chemical_classification Sequence Homology Amino Acid Tryptophan virus diseases RNA RNA-Binding Proteins General Medicine Transfection Phosphoproteins RGG Virology Molecular biology digestive system diseases Amino acid Protein Structure Tertiary chemistry HCV COS Cells Mutation Plasmids Protein Binding |
| Zdroj: | Journal of biochemistry. 141(6) |
| ISSN: | 0021-924X |
| Popis: | 金沢大学がん研究所 Hepatitis C Virus (HCV) non-structural proteins are major components of replication complex that is modulated by several host factors. We previously reported that nucleolin, a representative nucleolar marker, interacts with the NS5B through two separated sequences, amino acids (aa) 208-214 and 500-506, and that W208 in the former stretch is essential for both nucleolin-binding and HCV replication. Here we evaluated the role of the latter stretch aa 500-506 of WRHRARS in nucleolin-binding and HCV replication scanned by alanine-substituted clustered mutant (cm) or point mutant (pm). One tryptophan and three arginine residues in the sequence were found to be essential both for nucleolin-binding in vivo and HCV replication detected with a HCV subgenomic replicon transfected into Huh7 cells. NS5B-binding of nucleolin was further delineated by truncation and clustered mutants of nucleolin. Arginine-glycine-glycine (RGG) repeat in the Glycine arginine rich (GAR) domain were defined to be indispensable for NS5B-binding immunologically detected in in vivo and in vitro although short internal-truncations of RGG repeat are tolerable for NS5B-binding. These results indicate that nucleolin is a critical host factor for HCV replication through the direct interaction between W208 and several residues at the sequence, aa 500-505, of NS5B, and the long-turn motif including RGG repeat at nucleolin C-terminal. © 2007 The Japanese Biochemical Society. |
| Databáze: | OpenAIRE |
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