| ○種別 (必須): | □ | 学術論文 (審査論文)
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| ○言語 (必須): | □ | 英語
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| ○カテゴリ (推奨): |
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| ○学究種別 (推奨): |
| ○組織 (推奨): | 1. | 徳島大学.疾患酵素学研究センター (2007年4月1日〜2016年3月31日/->組織[徳島大学.先端酵素学研究所.次世代酵素学研究領域])
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| ○著者 (必須): | 1. | (英) Mine Akira (日) (読)
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| 2. | (英) Takeda Atsushi (日) (読)
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| 3. | 谷口 貴子
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| 4. | 谷口 寿章
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| 5. | (英) Kaido Masanori (日) (読)
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| 6. | (英) Mise Kazuyuki (日) (読)
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| 7. | (英) Okuno Tetsuro (日) (読)
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| ○題名 (必須): | □ | (英) Identification and characterization of the 480-kilodalton template-specific RNA-dependent RNA polymerase complex of red clover necrotic mosaic virus. (日)
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| ○要約 (任意): | □ | (英) Replication of positive-strand RNA viruses occurs through the assembly of membrane-associated viral RNA replication complexes that include viral replicase proteins, viral RNA templates, and host proteins. Red clover necrotic mosaic virus (RCNMV) is a positive-strand RNA plant virus with a genome consisting of RNA1 and RNA2. The two proteins encoded by RNA1, a 27-kDa protein (p27) and an 88-kDa protein containing an RNA-dependent RNA polymerase (RdRP) motif (p88), are essential for RCNMV RNA replication. To analyze RCNMV RNA replication complexes, we used blue-native polyacrylamide gel electrophoresis (BN/PAGE), which enabled us to analyze detergent-solubilized large membrane protein complexes. p27 and p88 formed a complex of 480 kDa in RCNMV-infected plants. As a result of sucrose gradient sedimentation, the 480-kDa complex cofractionated with both endogenous template-bound and exogenous template-dependent RdRP activities. The amount of the 480-kDa complex corresponded to the activity of exogenous template-dependent RdRP, which produced RNA fragments by specifically recognizing the 3'-terminal core promoter sequences of RCNMV RNAs, but did not correspond to the activity of endogenous template-bound RdRP, which produced genome-sized RNAs without the addition of RNA templates. These results suggest that the 480-kDa complex contributes to template-dependent RdRP activities. We subjected those RdRP complexes to affinity purification and analyzed their components using two-dimensional BN/sodium dodecyl sulfate-PAGE (BN/SDS-PAGE) and mass spectrometry. The 480-kDa complex contained p27, p88, and possible host proteins, and the original affinity-purified RdRP preparation contained HSP70, HSP90, and several ribosomal proteins that were not detected in the 480-kDa complex. A model for the formation of RCNMV RNA replication complexes is proposed. (日)
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| ○キーワード (推奨): | 1. | (英) Host-Parasite Interactions (日) (読)
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| 2. | (英) Macromolecular Substances (日) (読)
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| 3. | (英) Molecular Weight (日) (読)
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| 4. | (英) Plant Diseases (日) (読)
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| 5. | (英) RNA Replicase (日) (読)
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| 6. | (英) RNA, Viral (日) (読)
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| 7. | (英) Templates, Genetic (日) (読)
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| 8. | (英) Tombusviridae (日) (読)
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| 9. | (英) Viral Proteins (日) (読)
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| 10. | (英) Virus Replication (日) (読)
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| ○発行所 (推奨): |
| ○誌名 (必須): | □ | Journal of Virology ([アメリカ微生物学会])
(pISSN: 0022-538X, eISSN: 1098-5514)
| ○ISSN (任意): | □ | 1098-5514
ISSN: 0022-538X
(pISSN: 0022-538X, eISSN: 1098-5514) Title: Journal of virologyTitle(ISO): J VirolPublisher: American Society for Microbiology (NLM Catalog)
(Scopus)
(CrossRef)
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| ○巻 (必須): | □ | 84
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| ○号 (必須): | □ | 12
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| ○頁 (必須): | □ | 6070 6081
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| ○都市 (任意): |
| ○年月日 (必須): | □ | 西暦 2010年 4月 7日 (平成 22年 4月 7日)
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| ○URL (任意): |
| ○DOI (任意): | □ | 10.1128/JVI.00054-10 (→Scopusで検索)
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| ○PMID (任意): | □ | 20375154 (→Scopusで検索)
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| ○備考 (任意): | 1. | (英) Article.Affiliation: Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kitashirakawa, Kyoto 606-8502, Japan. (日)
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| 2. | (英) Article.PublicationTypeList.PublicationType: Journal Article (日)
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| 3. | (英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't (日)
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| 4. | (英) OtherID: PMC2876630 (日)
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