Regulation of RNA degradation by RNase

RNase 调节 RNA 降解

• 批准号: 17026001
• 负责人: NAITO Satoshi
• 金额: $ 14.72万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research on Priority Areas
• 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-17026001/
• 关键词: mRNA Degradation; translation arrest; colicin; cystathionine gamma-synthase; S-adenosylmethionine; シスタチオニンγ-シンターゼ; シスタチオニン γシンターゼ

When cystathionine gamma-synthase (CGS) mRNA of Arabidopsis is translated in vitro in the presence of S-adenosylmethionine (SAM), translation arrest is induced at Ser-94, which then induces CGS mRNA degradation. mRNA that carries the first exon coding region of CGS was prepared by in vitro translation and was used to program in vitro translation of wheat germ extract. Ribosome was stalled at the step of translocation and the peptidyl-tRNA, the translation intermediate, was located in the A-site. A series of mRNA degradation intermediates, whose 5' ends were separated by about 30 nucleotides from each other, were identified. Evidence was found that shows that these degradation intermediates correspond to ribosomes that were stacked behind the initial stalled ribosome. Poly (G) has been reported to inhibit RNase activity in the in vitro translation system of wheat germ extract. When Poly(G) was added to the translation reaction mixture, a 5'-fragment of CGS mRNA whose 3' end matches with one of the 5' -ends of the degradation intermediate. The result suggested that the mRNA degradation occurs by an endoribonucleolytic digestion.An Escherichia coli ribonuclease colicin E5 digests the anticodon loop of tRNAs for Trp, His, Asn and Asp. Colicin E5 preferentially digests GU dinucleotide. Structure and substrate specificity relationship of colicin E5 were studied. The substrate specificity could be explained by the structural constraints at the substrate pocket. Effects of expression of colicin D that digests tRNA(Arg) in budding yeast and HeLa cells were analyzed.

在s -腺苷甲硫氨酸（SAM）存在下，拟南芥胱硫氨酸γ -合成酶（CGS） mRNA在体外翻译时，在Ser-94位点诱导翻译阻滞，进而诱导CGS mRNA降解。通过体外翻译制备了携带CGS第一外显子编码区的mRNA，并将其用于小麦胚芽提取物的体外翻译编程。核糖体在易位阶段被阻滞，翻译中间体肽基trna位于a位点。鉴定出一系列mRNA降解中间体，其5'端彼此相距约30个核苷酸。发现的证据表明，这些降解中间体对应于堆积在初始停滞核糖体后面的核糖体。Poly (G)在小麦胚芽提取物的体外翻译系统中具有抑制RNase活性的作用。当Poly(G)加入到翻译反应混合物中时，CGS mRNA的一个5‘片段，其3’端与降解中间体的一个5'端匹配。结果表明，mRNA的降解是通过核糖核内溶解消化发生的。大肠杆菌核酸酶大肠杆菌素E5可消化trna的反密码子环，包括trna中的trna， trna中的trna。Colicin E5优先消化GU二核苷酸。研究了大肠杆菌素E5的结构与底物特异性关系。底物特异性可以用底物袋处的结构约束来解释。分析了消化tRNA（Arg）的大肠杆菌素D在出芽酵母和HeLa细胞中表达的影响。

项目成果

Construction of a positive selection marker by a lethal gene with the amber stop codon(s) regulator.

通过带有琥珀终止密码子调节器的致死基因构建正选择标记。

• 发表时间: 2006
• 期刊: Biosci. Biotech. Biochem. 70
• 作者: S. Yajima;T. Ogawa;K. Takahashi;S. Ohashi-Kumihiro;S. Ohashi-Kumihiro
• 通讯作者: S. Ohashi-Kumihiro

Esterification of Eschericia coli tRNAs with D-histidine and D-lisine by aminoacyl-tRNA synthetases.

通过氨酰基-tRNA 合成酶将大肠杆菌 tRNA 与 D-组氨酸和 D-赖氨酸酯化。

• 发表时间: 2005
• 期刊: Biosci. Biotech. Biochem. 69
• 作者: Tetsuhiro Ogawa;Sakura Inoue;Shunsuke Yajima;Makoto Hidaka;Haruhiko Masaki;K.Takahashi;S.Ohashi-Kumihiro;T. Takayama
• 通讯作者: T. Takayama

Nascent peptide-mediated translation elongation arrest coupled with mRNAdegradation in the CGS1 gene of Arabidopsis.

拟南芥 CGS1 基因中新生肽介导的翻译延伸停滞与 mRNA 降解相结合。

• 发表时间: 2005
• 期刊: Genes Dev. 19
• 作者: Tetsuhiro Ogawa;Sakura Inoue;Shunsuke Yajima;Makoto Hidaka;Haruhiko Masaki;K.Takahashi;S.Ohashi-Kumihiro;T. Takayama;H. Onouchi
• 通讯作者: H. Onouchi

Sequence-specific recognition of colicin E5, a tRNA-targeting ribonuclease.

• DOI: 10.1093/nar/gkl629
• 发表时间: 2006
• 期刊: Nucleic acids research
• 影响因子: 14.9
• 作者: Ogawa T;Inoue S;Yajima S;Hidaka M;Masaki H
• 通讯作者: Masaki H

CGS遺伝子発現の転写後自己制御機構 : 翻訳を停止したリボソームの解析

CGS基因表达的转录后自动调节机制：停止翻译的核糖体分析

• 发表时间: 2007
• 作者: Tetsuhiro Ogawa;Sakura Inoue;Shunsuke Yajima;Makoto Hidaka;Haruhiko Masaki;K.Takahashi;S.Ohashi-Kumihiro;T. Takayama;H. Onouchi;T. Takayama;H. Onouchi;H.Onouchi;T.Takayama;尾之内均;原口雄飛;尾上典之;川崎大輔;門倉嘉知
• 通讯作者: 門倉嘉知