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Molecular genetic elucidation of the function of GTP binding protein to be concerned with the membrane phospholipids in Escherichia coli

大肠杆菌GTP结合蛋白与膜磷脂相关功能的分子遗传学阐明

基本信息

批准号：
15580057
负责人：
MATSUZAKI Hiroshi
金额：
$ 2.24万
依托单位：
Saitama university
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

项目摘要

This studies aim the elucidation of the function of GTP binding protein (G protein) to be concerned with the function of the membrane phospholipids in E.coli. The influence on the transcription or translation activities in the membrane phospholipids deficient mutation etc. stock was examined by using a promoter strength measurement system that made strong fluorescence GFP as a reporter gene and a real-time RT-PCR method about low molecular weight G protein gene yihA, era, and obgE essential on growth in E..coli. A clear difference was not admitted in the cardiolipin deficient mutation and the transcription activity of low molecular weight G protein gene increased remarkably in the acidic phospholipids deficient mutation (pgsA3 mutation stock) compared with the wild-type strain. Thus, it was suggested that the membrane phospholipid composition, especially the increase and decrease of the acidic pospholipid content take part in the appearance control.【summary of research and view】G protein gene, yihA gene of an essential G protein of E.coli were admitted the increase of the transcript revitalization in the acid phosphorus lipid synthesis loss mutation stock, and clarified that this expression depended on the expression of the acidic phospholipid synthesis enzyme gene. Der (or Yfg) related to presumption G protein localized in the septum in a cell by binding with GFP in the complete amphiphilic phospholipid defincient mutation. The above-mentioned result depends on the amount of the acidic phospholipid by the expression of G protein and suggests G protein, the membrane lipid, and the possibility of the interaction with the biomembrane strongly.

本研究旨在阐明大肠杆菌GTP结合蛋白（G蛋白）的功能与膜磷脂的功能有关。利用以强荧光GFP为报告基因的启动子强度测定系统和大肠杆菌生长所必需的低分子量G蛋白基因yihA、era和obgE的实时荧光定量PCR方法，研究了膜磷脂缺陷突变等对转录或翻译活性的影响。杆菌与野生型菌株相比，心磷脂缺陷突变没有明显差异，酸性磷脂缺陷突变（pgsA3突变株）中低分子量G蛋白基因的转录活性显着增加。因此，认为膜磷脂组成，特别是酸性磷脂含量的增加和减少参与了外观控制。[研究概要和观点]大肠杆菌必需G蛋白的G蛋白基因、yihA基因在酸性磷脂质合成缺失突变株中的转录物再生增加，并阐明了该表达依赖于酸性磷脂合成酶基因的表达。Der（或Yfg）与推测G蛋白相关，通过与GFP结合定位于细胞间隔中的完全两亲性磷脂明确突变。上述结果取决于G蛋白表达的酸性磷脂的量，并提示G蛋白、膜脂质和与生物膜强烈相互作用的可能性。