| Project/Area Number |
11694257
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Pathological medical chemistry
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| Research Institution | KANAZAWA UNIVERSITY |
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Principal Investigator |
MURAKAMI Seishi Dept.Mol.Oncology, Cancer Res.Inst., KANAZAWA UNIVERSITY Professor, がん研究所, 教授 (90019878)
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| Co-Investigator(Kenkyū-buntansha) |
HAYASHI Naoyuki Dept.Mol.Oncology, Cancer Res.Inst., KANAZAWA UNIVERSITY Research Associate, がん研究所, 助手 (50253456)
YOSHIOKA Katsuji Dept.Mol.Cell Biol., Cancer Res.Inst., KANAZAWA UNIVERSITY Associate Professor, がん研究所, 助教授 (60200937)
SUDA Takashi Center for Mol.Target Drugs Cancer Res.Inst., KANAZAWA UNIVERSITY Professor, がん研究所, 教授 (70250090)
NOMURA Takahiro Dept.Mol.Oncology, Cancer Res.Inst., KANAZAWA UNIVERSITY Research Associate, がん研究所, 助手 (80115261)
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| Project Period (FY) |
1999 – 2000
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| Keywords | HCV NS5B / RNA-dependent RNA polymerase(RdRP) / alanine scanning / RNA binding / HCV infection / activation of NF-kB |
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| Research Abstract |
Eradication of HCV has been expected to reduce or prevent incidence of HCC.HCV NS5B, RNA-dependent RNA polymerase(RdRP), is a target to inhibit HCV replication. We constructed a series of clustered and point alanine substitution mutations of NS5B and examined the effects of the mutations on functions of NS5B.The results are followings. 1)5 amino acid residues were newly identified to be indispensable for RdRP activity(Hepatology, in press). All are outside of the motifs conserved among RdRPs and reverse transcriptases. The result may provide desigins for specific inhibitors for HCV RdRP.2)RdRP activity of NS5B requires not template binding but template/primer. All mutants negative in the template binding are positive in RdRP activity except Y276A which is both negative in template/primer binding and RdRP activity. 3)The two residues apart from the catalytic center of RdRP activity are important for oligomerization of NS5B which we could find to be essential for RdRP activity. 4)Overexpression of NS5B activated AP-1 and NF-kB critical for acute inflammatory response, but a NS5B sequence could not be specified to be responsible for the activation which is influenced by expression level of NS5B.NS5B may modulate inflammatory responses but it remains addressed whether our observation has biological relevance in chronic HCV infection where a low amount of NS5B with coordinated expression of the other NS proteins is expected.
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