YEAST HISTONE ACETYLATION GENES AND MUTANTS

描述：本研究的长期目标是了解 组蛋白乙酰化在染色质结构和功能，使用酵母 S.酿酒厂来研究这个过程。 由于大多数酵母蛋白质似乎 有人类同源物，这个项目应该揭示人类生物学， 疾病 具体目的是：1）确定酵母的功能 组蛋白H4乙酰转移酶HAT 1基因已被鉴定并克隆 去年 由于hat 1无效突变体没有明显的表型， 需要方法来确定HAT 1蛋白的体内功能。 2)为了表征HAT 1相互作用蛋白，HIF 1和HIF 2， 两个混合动力系统。 假设HIF 2和其他相关的 蛋白质，靶酶如组蛋白乙酰转移酶和组蛋白 将测试脱乙酰酶对组蛋白的作用。 3)寻找其他组蛋白 乙酰转移酶突变体，使用与所用方法相似的筛选方法 获得hat 1 -1突变体。 对应于N-末端尾部的肽 四个核心组蛋白中的每一个将被用作底物， 乙酰转移酶测定，筛选酵母菌群的提取物 温度敏感（TS）突变体，以找到缺陷， 乙酰转移酶活性 一旦发现突变体， 基因将被克隆，基因产物的作用将被确定。 4)到 检查其他几种酵母基因产物，看它们是否是组蛋白 修饰酶

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.