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Molecular basis of dihydrouridine formation on tRNA

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タイトル: Molecular basis of dihydrouridine formation on tRNA
著者: Yu, Futao 著作を一覧する
Tanaka, Yoshikazu 著作を一覧する
Yamashita, Keitaro 著作を一覧する
Suzuki, Takeo 著作を一覧する
Nakamura, Akiyoshi 著作を一覧する
Hirano, Nagisa 著作を一覧する
Suzuki, Tsutomu 著作を一覧する
Yao, Min 著作を一覧する
Tanaka, Isao 著作を一覧する
キーワード: protein-tRNA complex
RNA modification
substrate recognition
X-ray crystallography
発行日: 2011年12月 6日
出版者: National Academy of Sciences
誌名: Proceedings of the National Academy of Sciences of the United States of America
巻: 108
号: 49
開始ページ: 19593
終了ページ: 19598
出版社 DOI: 10.1073/pnas.1112352108
抄録: Dihydrouridine (D) is a highly conserved modified base found in tRNAs from all domains of life. Dihydrouridine synthase (Dus) catalyzes the D formation of tRNA through reduction of uracil base with flavin mononucleotide (FMN) as a cofactor. Here, we report the crystal structures of Thermus thermophilus Dus (TthDus), which is responsible for D formation at positions 20 and 20a, in complex with tRNA and with a short fragment of tRNA (D-loop). Dus interacts extensively with the D-arm and recognizes the elbow region composed of the kissing loop interaction between T- and D-loops in tRNA, pulling U20 into the catalytic center for reduction. Although distortion of the D-loop structure was observed upon binding of Dus to tRNA, the canonical D-loop/T-loop interaction was maintained. These results were consistent with the observation that Dus preferentially recognizes modified rather than unmodified tRNAs, indicating that Dus introduces D20 by monitoring the complete L-shaped structure of tRNAs. In the active site, U20 is stacked on the isoalloxazine ring of FMN, and C5 of the U20 uracil ring is covalently cross-linked to the thiol group of Cys93, implying a catalytic mechanism of D20 formation. In addition, the involvement of a cofactor molecule in uracil ring recognition was proposed. Based on a series of mutation analyses, we propose a molecular basis of tRNA recognition and D formation catalyzed by Dus.
資料タイプ: article (author version)
出現コレクション:雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

提供者: 田中 良和


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