Molecular Evolution of RNA modification enzymes and RNA recognition mechanism

RNA修饰酶的分子进化和RNA识别机制

• 批准号: 17613003
• 负责人: HORI Hiroyuki
• 金额: $ 2.11万
• 依托单位: Ehime University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17613003/
• 关键词: Nucleic Acid; Enzyme; Genome; Evolution; Biomolecule; RNA修飾; 好熱菌; tRNA; RNAメチル化酵素

In this project, we have obtained many findings in the RNA modification field.Main findings are as follows.(1) Construction of in vitro RNA modification system under high temperaturesWe have clarified roles of thermophilic-bacteria specific polyamines against the RNA modification machinery.(2) Clarifications of RNA recognition mechanisms of RNA modification enzymesWe have reported that Aquifex aeolicus TrmD [tRNA (m1 G37) methyltransferase] has a unique RNA recognition mechanism.The enzyme methylates tRNA molecules possessing not only G36G37 sequence but also A36G37 sequence. Furthermore, the enzyme methylates micro-helix RNA possessing A36G37 sequence. These results showed that A. aeolicus TrmD has a broad substarte specificity in comparison with other TrmD proteins.We have reported that eukaryote tRNA (m7G46) methyltransferase [Trm8/Trm82 complex] recognizes the D-arm structure in tRNA, demonstrating that eukaryote enzyme has a strict RNA recognition mechanism as compared to bacterial enzymes.(3) Clarifications of relationship between protein structure and RNA recognition mechanismWe performed site-directed mutagenesis study against basic amino acid residues conserved in TrmH [tRNA (Gm18) methyltransferases]. We could distinguish residues required in the first tRNA binding process and residues required in the progress of the catalytic cycles.

在本项目中，我们在RNA修饰领域取得了许多成果。主要发现如下：(1)高温下体外RNA修饰体系的构建我们阐明了嗜热菌特异性多胺对RNA修饰机制的作用。(2)RNA修饰酶的RNA识别机制我们已经报道了Aquifex aeolicus TrmD[tRNA(M1 G37)甲基转移酶]具有独特的RNA识别机制。该酶甲基化tRNA分子不仅具有G36G37序列，还具有A36G37序列。此外，该酶还甲基化了具有A36G37序列的微螺旋RNA。我们已经报道了真核生物tRNA(M7G46)甲基转移酶[Trm8/Trm82复合体]识别tRNA中的D-臂结构，表明与细菌酶相比，真核生物酶具有严格的RNA识别机制。(3)蛋白质结构与RNA识别机制之间关系的阐明我们对TrmH[tRNA(Gm18)甲基转移酶]中保守的碱性氨基酸残基进行了定点突变研究。我们可以区分第一次tRNA结合过程中所需的残基和催化循环过程中所需的残基。

项目成果

Detection of structural changes in a cofactor binding protein by using a wheat germ cell-free protein synthesis system coupled with unnatural amino acid probing

• DOI: 10.1002/prot.21341
• 发表时间: 2007-05-15
• 期刊: PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
• 影响因子: 2.9
• 作者: Abe, Masato;Ohno, Satoshi;Endo, Yaeta
• 通讯作者: Endo, Yaeta

The core domain of Aquifex aeolicus tRNA (m^7G46) methyltransferase has the methyl-transferactivity to tRNA

Aquifex aeolicus tRNA (m^7G46) 甲基转移酶的核心结构域具有对 tRNA 的甲基转移活性

• 发表时间: 2006
• 期刊: Nucleic Scids Res. Symp. Ser. 50
• 作者: Hee-Jin Kang;Koichi Uegaki;Harumi Fukada;Kazuhiko Ishikawa;C.Tomikawa
• 通讯作者: C.Tomikawa

A novel tRNA recognition mechanism of Aquifex aeolicus Trm1 [tRNA (<m^2>_2G26)methyltransferase]

Aquifex aeolicus Trm1 的一种新型 tRNA 识别机制 [tRNA (<m^2>_2G26)methyltransferase]

• 发表时间: 2005
• 期刊: Nucleic Acids Res. Symp. Ser. 49
• 作者: ISOZAKI;Hatsuhito;T.Awai
• 通讯作者: T.Awai

Structural change of tRNA (Gm18) methyltransferase by binding of methyl donor analogues.

通过甲基供体类似物的结合来改变 tRNA (Gm18) 甲基转移酶的结构。

• 发表时间: 2005
• 期刊: Nucleic Acids Res. Symp. Ser. 49
• 作者: Nunomura S;Gon Y;Yoshimaru T;Suzuki Y;Nishimoto H;Kawakami T;Ra C.;Koga K.et al.;SUGIHARA;K.Watanabe
• 通讯作者: K.Watanabe

A novel tRNA recognition mechanism of Aquifex aeolicus Trm1 [tRNA (m^2_2G26) methyltransferase].

Aquifex aeolicus Trm1 的一种新型 tRNA 识别机制 [tRNA (m^2_2G26) 甲基转移酶]。

• 发表时间: 2005
• 期刊: Nucleic Acids Res. Symp. Ser. 49
• 作者: Siddiqui;M. A;Kaoru Nakasone;Katsuya Satoh et al.;T.Awai
• 通讯作者: T.Awai