YEAST HISTONE ACETYLATION GENES AND MUTANTS

描述：这项研究的长期目标是了解 组蛋白乙酰化对染色质结构和功能的影响 酿酒酵母研究这一过程。因为大多数酵母蛋白似乎 有人类的同源物，这个项目应该会揭示人类生物学和 疾病。具体目标是：1)确定酵母的功能 组蛋白H4乙酰转移酶，HAT1，其基因被鉴定和克隆 去年。由于HAT1缺失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)鉴定HAT1相互作用蛋白HIF1和HIF2 使用双混合系统。假设HIF2和其他相关的 蛋白质，靶向酶，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 使用与所用程序类似的筛选程序筛选乙酰转移酶突变体 以获得HAT1-1突变体。与N-末端尾部对应的多肽 四个核心组蛋白的每一个都将被用作底物 乙酰转移酶分析，筛选酵母集合的提取物 温度敏感型(Ts)突变体发现有缺陷的 乙酰转移酶活性。一旦确定了突变体，相应的 基因将被克隆，基因产物的作用将被确定。4)至 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。

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.