YEAST HISTONE ACETYLATION GENES AND MUTANTS

酵母组蛋白乙酰化基因和突变体

• 批准号: 2701819
• 负责人: Rolf Sternglanz
• 金额: $ 17.92万
• 依托单位: STATE UNIVERSITY NEW YORK STONY BROOK
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 2001-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2701819
• 关键词: Saccharomyces cerevisiae; acetylation; acyltransferase; chromatin; enzyme activity; fungal genetics; gene mutation; histones; molecular cloning; protein structure function; temperature sensitive mutant

DESCRIPTION: The long term goal of this research is to understand the role of histone acetylation in chromatin structure and function, using the yeast S. cerevisiae to study the process. Since most yeast proteins appear to have human homologs, this project should shed light on human biology and disease. The specific aims are: 1) to determine the function of the yeast histone H4 acetyltransferase, HAT1, whose gene was identified and cloned last year. Since a hat1 null mutant has no obvious phenotypes, other approaches are needed to determine the in vivo function of the HAT1 protein. 2) to characterize the HAT1-interacting proteins, HIF1 and HIF2, identified with the two-hybrid system. The hypothesis that HIF2, and other related proteins, target enzymes such as histone acetyltransferases and histone deacetylases to the histones will be tested. 3) to find other histone acetyltransferase mutants using screening procedures similar to the one used to obtain the hat1-1 mutant. Peptides corresponding to the N-terminal tails of each of the four core histones will be used as substrates for acetyltransferase assays, screening extracts from a collection of yeast temperature-sensitive (ts) mutants to find ones defective in acetyltransferase activity. Once mutants are identified, the corresponding genes will be cloned and the roles of the gene products determined. 4) to examine several other yeast gene products to see if they are histone modifying enzymes.

描述：这项研究的长期目标是了解其作用 使用酵母研究染色质结构和功能中的组蛋白乙酰化 酿酒酵母来研究这个过程。 由于大多数酵母蛋白似乎 具有人类同源物，该项目应该揭示人类生物学和 疾病。 具体目的是：1）确定酵母的功能 组蛋白H4乙酰转移酶HAT1基因被鉴定并克隆 去年。 由于 hat1 无效突变体没有明显的表型，其他 需要一些方法来确定 HAT1 蛋白的体内功能。 2) 表征 HAT1 相互作用蛋白 HIF1 和 HIF2，已鉴定 与双混合系统。 HIF2 和其他相关的假设 蛋白质、靶酶，例如组蛋白乙酰转移酶和组蛋白 将测试组蛋白的脱乙酰酶。 3）寻找其他组蛋白 使用与所使用的类似的筛选程序来筛选乙酰转移酶突变体 获得 hat1-1 突变体。 N-末端尾部对应的肽 四个核心组蛋白中的每一个都将用作底物 乙酰转移酶测定，筛选酵母集合中的提取物 温度敏感（ts）突变体以发现缺陷 乙酰转移酶活性。 一旦突变体被识别，相应的 基因将被克隆并确定基因产物的作用。 4）到 检查其他几种酵母基因产物，看看它们是否是组蛋白 修饰酶。