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REGULATION OF RETROTRANSPOSITION IN S CEREVISIAE

酿酒酵母逆转录转座的调控

基本信息

批准号：
6019046
负责人：
M Joan CURCIO
金额：
$ 11.21万
依托单位：
WADSWORTH CENTER
依托单位国家：
美国
项目类别：
财政年份：
1995
资助国家：
美国
起止时间：
1995-08-01 至 2000-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6019046
关键词：
Saccharomyces cerevisiae endopeptidases enzyme induction /repression fungal genetics gene complementation gene expression gene mutation genetic transcription reporter genes transposon /insertion element

项目摘要

DESCRIPTION: The Ty1 elements of yeast are retrotransposons that encode structural proteins, protease, integrase, and reverse transcriptase. Like retroviruses, these products are formed by transcription of polycistronic messages and post-translational processing events such as protease cleavages and phosphorylation. Although Ty1 RNA represents a large fraction of total RNA in the cell, transposition events are rare, presumably due to a mechanism of regulation called transpositional dormancy. The main objective of this proposal is to identify and characterize the cellular genes involved in transpositional dormancy. These genes are identified by mutations that cause a hypertranspositional phenotype. The PI has used a novel reporter gene construct to screen for such mutations and at present over 160 mutations have been isolated. The initial focus will be on characterizing a subset of these which affect transposition at a post-transcriptional level rather than those mutations that increase Ty1 transcription rates. Genetic characterization of the mutants will include complementation test, dominance relationships, and several genetic and molecular screens to determine the level at which the mutation affects the transposition process. Selected genes will be cloned by complementation of the hypertranspositional phenotype. Steps in the formation of Ty1-like particles, such as protease activation (a rate-limiting step) and VLP formation will be studied in mutant and wild- type cells to identify new mechanisms of transpositional regulation.

描述：酵母的Ty 1元件是逆转录转座子，结构蛋白、蛋白酶、整合酶和逆转录酶。像逆转录病毒一样，这些产物是通过转录多顺反子信息和翻译后加工事件，蛋白酶裂解和磷酸化。虽然Ty 1 RNA代表一种细胞中总RNA的大部分，转座事件是罕见的，这大概是由于一种叫做转座的调节机制休眠本提案的主要目标是确定和描述参与易位休眠的细胞基因。这些基因通过引起高转座表型的突变来鉴定。的 PI使用了一种新的报告基因构建体来筛选此类突变目前已经分离出超过160种突变。最初的重点将是对这些影响转置的子集的特征在转录后水平，而不是那些增加 Ty 1转录速率。突变体的遗传特征将包括互补性测试、显性关系以及一些基因和分子筛选，以确定突变影响转座过程。选定的基因将通过高转座表型的互补克隆。步骤在Ty 1样颗粒的形成中，例如蛋白酶活化（a 限速步骤）和VLP形成将在突变体和野生型中进行研究- 类型的细胞，以确定新的机制的易位调节。