Ash1p Mediated Transcriptional Repression

Ash1p 介导的转录抑制

• 批准号: 6640473
• 负责人: LENNY M CARRUTHERS
• 金额: $ 4.64万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6640473
• 关键词: Saccharomyces cerevisiae; chromatin; fungal proteins; gene induction /repression; genetic screening; immunoprecipitation; nucleoproteins; protein protein interaction; protein structure function; transcription factor; yeast two hybrid system

DESCRIPTION (provided by applicant): Ash1p is a nuclear protein found in budding yeast that is an intrinsic factor responsible for generating cell diversity. An example of this process is asymmetric cell division, a significant developmental event that occurs in organisms ranging from bacteria, yeast, worms, and mammals. A major function of Ash1p in Saccharomyces cerevisiae is that of a transcriptional repressor. This protein is preferentially localized to the daughter nucleus following nuclear division. Ash1p actively inhibits transcription of the gene encoding for the HO endonuclease that, in turn, is responsible for mating type switching.While many details have been defined for the mechanism leading to the asymmetric localization of Ash1p, lithe has been learned about its function as a transcriptional repressor. Therefore, the overall objective of the proposal is to understand both the regulation and mechanism of transcriptional repression by Ash1p. To address these goals the first aim of this proposal will utilize yeast molecular and classical genetics to search for proteins that may be required for Ash1p function, along with the yeast two-hybrid system to probe for other proteins that may interact with Ash1p. Aim 2 will utilize chromatin immunoprecipitation assays to define the environment and components that exist at the HO gene when it is being repressed. And aim 3 will examine the chromatin structure of the HO locus in the presence of Ash1p.

描述（由申请人提供）： ASH1P是在发芽酵母中发现的一种核蛋白，是负责产生细胞多样性的内在因素。这个过程的一个例子是不对称的细胞分裂，这是一个重要的发育事件，发生在细菌，酵母，蠕虫和哺乳动物的生物中。 ASH1P在酿酒酵母中的主要功能是转录阻遏物。该蛋白在核分裂后优先将其定位在子核上。 ASH1P积极抑制针对HO内核酸酶的基因的转录，该基因反过来又负责交配类型的转换。尽管已定义了许多细节用于导致ASH1P不对称定位的机制，但Lithe已被学到了其作为转录抑制器的功能。因此，该提案的总体目标是了解ASH1P的转录抑制的调节和机制。为了解决这些目标，该提案的第一个目的将利用酵母分子和经典的遗传学来搜索ASH1P功能可能需要的蛋白质，以及酵母双杂交系统，以探测其他可能与ASH1P相互作用的蛋白质。 AIM 2将利用染色质免疫沉淀测定法来定义HO基因被抑制时存在的环境和成分。 AIM 3将在ASH1P存在下检查HO基因座的染色质结构。