拟南芥MTERF家族蛋白调控叶绿体基因转录终止的分子机制

项目名称： 拟南芥MTERF家族蛋白调控叶绿体基因转录终止的分子机制

项目编号： No.31470337

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 生物科学

项目作者： 崔永兰

作者单位： 上海师范大学

项目金额： 80万元

中文摘要： 叶绿体基因表达对光合作用及植物发育至关重要。动物线粒体转录终止因子（MTERF）家族是一类多功能蛋白，参与线粒体基因转录终止等过程。拟南芥MTERF家族含有35个成员，其中12个定位于叶绿体，包括PDE191，其敲除突变体呈白化致死表型，叶绿体rpoA及petD等基因转录终止异常。本项目我们将深入研究拟南芥PDE191以及其他叶绿体定位的MTERF家族蛋白的功能。具体研究内容包括: 1）利用CHIP结合EMSA技术分析PDE191与rpoA等叶绿体基因的体内外结合情况，分离与PDE191互作蛋白进一步分析PDE191调控rpoA等基因转录终止的机制；2）通过分析rpoA表达及PEP活性阐明PDE191突变导致植株白化致死原因；3）分析拟南芥叶绿体定位的MTERF家族成员各自调控的下游基因，揭示叶绿体基因转录终止调控的一般过程。该项目的完成将促进我们对叶绿体基因转录终止机制的深入理解。

中文关键词： 拟南芥；叶绿体发育；转录终止；分子机制

英文摘要： Chloroplast gene expression is crucial for photosynthesis and plant development. The mitochondrial transcription termination factor (MTERF) is a multifunction protein family, which is involved in transcription termination of mitochondrial genes. Arabidopsis contains at least 35 genes for MTERF family proteins. Of 35 MTERF proteins, 12 are plastid-localized, including the PDE191 gene, whose knocked-out mutant is albino and seedling lethal, with abnormal transcription termination of chloroplast genes,including rpoA and petD. In the present project, we will further study the PDE191 gene function and systematic analysis of the MTERF proteins regulating the transcription termination of chloroplast genes, which includes: 1) analyzing the binding sites of PDE191 on the rpoA gene with CHIP (chromosome immunoprecipitation) and EMSA((electrophoresis mobility shift assay) methods, characteristication of interacting proteins with PDE191 protein; 2) elucidating the causes of albino lethal phenotype of pde191 by rpoA expression and PEP activity analysis; 3) Identification of the downstream target genes of plastid-localized MTERF proteins in Arabidopsis, revealing the general process of transcription termination of chloroplast genes. The completion of the project will facilitate our understanding of the molecular mechanism underlying transcription termination of chloroplast genes.

英文关键词： Arabidopsis;chloroplast development;transcription termination;molecular mechanism