Systematic analyses of biological functions of helicase-like proteins in eukaryote

真核生物解旋酶样蛋白生物学功能的系统分析

基本信息

批准号：
11672202
负责人：
EKI Toshihiko
金额：
$ 1.02万
依托单位：
RIKEN
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672202/
关键词：
helicase family genome functional study budding yeast nematode protein-protein interaction RNAi gene disruption expression profile 遺伝子機能破壊出芽酵母ゲノムヘリカーゼ様蛋白質線虫ゲノム RNAヘリカーゼ YDL084W遺伝子 pre-mRNA splicing 核酸依存性ATPase

项目摘要

This research has aimed to identify all members of helicase-like proteins in eukaryote and clarify their biological functions. I have focused on two scientific subjects described below.1. Functional study of novel helicase-like proteins that were found by bioinformatics.2. Functional study of nematode helicase-like proteins by characterizing RNAi mutants.I have newly obtained the following observations throughout this research.1. Six out of 20 novel ORFs that were found in budding yeast, were identified to be essential by gene disruptions.2. Several genes that were highly expressed under specific conditions were identified by Northern analyses of 43 helicase-like ORFs. It also clarified that the levels of transcripts from more than 70% of examined helicase-like genes significantly increased upon heat-shock.3. Ydl084wp has been newly found to be RNA-dependent ATPase.4. Interactions of gene products such as Yhl218wp and YklO 17cp with several cellular proteins were detected by yeast two-hybrid assays using 10 novel nonessenfial proteins. Some genetic evidences suggested that Ynl218wp might act in DNA recombination.5. It has been clarified that three novel gene products (i.e., Ydr332wp, Ygl064cp, and Yol095cp) play important roles in mitochondria functions.6. Systematic characterization of nematode RNAi mutants of helicase-like genes revealed the evolutional conservation of the genes and their functions in eukaryote. Since more than 40% of examined RNAi mutants exhibited various kinds of abnormalities in cell. proliferation and development, suggesting biological importance of helicase-like proteins in higher eukaryote.

这项研究旨在鉴定真核生物中所有类似解旋酶的蛋白质成员，并阐明其生物学功能。我专注于下面描述的两个科学主题1。生物信息学发现的新型解旋酶样蛋白的功能研究2。通过表征RNAi突变体来对线虫解旋酶样蛋白的功能研究。i在整个研究中新获得了以下观察结果。1。在萌芽的酵母中发现的20个新型ORF中，有6个被基因破坏确定为必不可少的。2。在特定条件下高度表达的几个基因通过43种解旋酶样ORF的北部分析鉴定出来。它还澄清说，来自70％以上检查的解旋酶样基因的转录本水平在热震中显着增加。3。 YDL084WP已被新发现是RNA依赖性ATPase.4。使用10种新型的非杂种蛋白检测到基因产物（例如YHL218WP和YKLO 17CP）与几种细胞蛋白的相互作用。一些遗传证据表明，YNL218WP可能作用于DNA重组5。已经澄清说，三种新型基因产物（即YDR332WP，YGL064CP和YOL095CP）在线粒体功能中起重要作用。6。类似于解酶样基因的线虫RNAi突变体的系统表征揭示了该基因及其在真核生物中的功能的进化保守性。由于超过40％的检查RNAi突变体在细胞中表现出各种异常。增殖和发育，表明在高级真核生物中类似解旋酶样蛋白的生物学重要性。