Gene Regulatory Codes and Signal/Regulatory Element Interactions in IME2

描述(申请人提供)：Honigberg实验室的长期目标是确定酵母调节其进入减数分裂途径的机制。这个系统提供了一个独特的机会来研究不同信号被整合以控制细胞命运的基本机制。该提案的第一个具体目标是确定IME2的基因调控密码，即确定控制该基因表达的所有调控元件以及它们对信号的反应。这一目标将通过对基因组IME2-LacZ等位基因的启动子进行广泛的突变来实现，然后在调节该基因的三种主要信号的所有组合下对每个IME2-x-LacZ突变进行分析。对这种序列和表达数据组合的分析将揭示基因调控密码背后的逻辑电路。该提案的第二个目标是确定IME2监管要素之间的相互作用。这一目标的第一部分将通过测量选定的IME2-x-LacZ等位基因对Ime1p和Rpd3p与IME2的URS1调控元件的关联的影响来实现。这一目标的第二部分将确定具有共同功能的IME2调控元件之间的相互作用。这将通过上位性(双突变分析)来实现。由于拟议的研究将导致对真核基因调控密码的第一次完整描述，完成这一项目为理解其他基因的调控密码提供了一个框架，包括其表达改变导致人类疾病的基因。转录调控缺陷对人类健康有着深远的影响，可影响糖尿病、癌症和帕金森氏症等多种疾病。由于转录调控的机制复杂且知之甚少，研究模式遗传有机体--发芽酵母酿酒酵母的转录调控可以带来一些基础性的发现，促进我们对这些疾病的病因学的理解。

DESCRIPTION (provided by applicant): The long-term objective of the Honigberg lab is to identify the mechanisms by which yeast regulate their transition into the meiotic pathway. This system presents a unique opportunity to study fundamental mechanisms by which diverse signals are integrated to control cellular fates. The first specific aim of the proposal is to determine the gene regulatory code for IME2, i.e. to identify all of the regulatory elements that control expression of this gene and the signal or signals to which they respond. This aim will be achieved through extensive mutagenesis of the promoter of a genomic ime2-lacZ allele, followed by assaying each ime2-x-lacZ mutant under all combinations of the three major signals that regulate this gene. Analysis of this combination of sequence and expression data will reveal the logic circuits underlying the gene regulatory code. The second aim of the proposal is to identify interactions between IME2 regulatory elements. The first part of this aim will be accomplished by measuring the effect of selected ime2-x-lacZ alleles on association of Ime1p and Rpd3p with the URS1 regulatory element of IME2. The second part of this aim will be to determine the interactions between IME2 regulatory elements that have shared functions. This will be accomplished by epistasis (double mutant analysis). Because the proposed research will result in the first complete description of a eukaryotic gene regulatory code, accomplishing this project provides a framework for understanding regulatory codes in other genes, including genes whose altered expression results in human disease. Defects in the regulation of transcription have profound effects on human health, affecting such diverse diseases as diabetes, cancer, and Parkinson's. Because the mechanism of transcriptional regulation is complex and still poorly understood, studying transcriptional regulation in a model genetic organism, the budding yeast S. cerevisiae, can lead to fundamental discoveries that advance our understanding of the etiology of these diseases.