| Project/Area Number |
11470080
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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 |
Virology
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| Research Institution | Kitasato University |
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Principal Investigator |
MIZUMOTO Kiyohisa Kitasato University, School of Pharmaceutical Sciences, Professor, 薬学部, 教授 (80092344)
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| Co-Investigator(Kenkyū-buntansha) |
OGINO Tomoaki University of Tokyo, Graduate School Researcher, 医学系研究科, 研究員 (80312213)
IWAMA Minako Kitasato University, School of Pharmaceutical Sciences, Research Associate, 薬学部, 助手 (10223421)
TSUKAMOTO Toshihiko Kitasato University, School of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (10236862)
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| Project Period (FY) |
1999 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥12,300,000 (Direct Cost: ¥12,300,000)
Fiscal Year 2002: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2001: ¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2000: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥3,800,000 (Direct Cost: ¥3,800,000)
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| Keywords | Paramyxovirus / Sendai virus / negative-strand RNA genome / RNA-dependent RNA polymerase / host factor / transcription / mRNA cap / tubulin / ウイルスmRNA合成 / mRNAキャップ構造 / キャップメチル化酵素 / ウイルスLタンパク質 / マイナス鎮RNAゲノム / ウイルスmRNA / mRNAキャッピング / パラミフソウイルス / パラミクソウイルス科 / in vitro転写反応 |
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| Research Abstract |
Sendai virus (SeV), a member of Paramyxovirus family, contains a monopartite negative strand RNA genome of approximately 15 kilobases. To elucidate the mechanism of transcription and replication of SeV, we developed an efficient and faithful in vitro transcription system using purified virus particles. We have previously shown that in vitro mRNA synthesis of SeV is almost entirely dependent on the addition of cellular proteins (host factors). In the present study, we further furified and characterized the host factor activity, and obtained the following results. 1) We purified the host factor activity, which stimulates in vitro SeV mRNA synthesis, from bovine brain, and found that the activity consists of four complementary protein factors, tubulin, phosphoglycerate kinase, enolase, and unidentified p24. 2) We showed that tubulin is involved in a transcription initiation complex formation by removing the viral M protein from the RNP, and by being integrated itself into the complex. On the other hand, phosphoglycerate kinase, enolase, and p24 are found to act at RNA shain elongation step. Furthermore, 3) we demonstrated that a recombinant viral L protein exhibits the cap methylase (mRNA guanine-7-methyltransferase) activity.
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