Analysis of MYST Family Acetyltransferase Functions

MYST家族乙酰转移酶功能分析

• 批准号: 7090639
• 负责人: LORRAINE PILLUS
• 金额: $ 28.67万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7090639
• 关键词: Saccharomyces; acetylation; acute myelogenous leukemia; acyltransferase; chromatin; enzyme activity; fungal genetics; gene induction /repression; gene mutation; genetic transcription; human genetic material tag; laboratory rabbit; molecular genetics; yeast two hybrid system

DESCRIPTION (provided by applicant): The long-term goal of this research is to understand mechanisms by which chromatin modifications regulate gene expression and chromosomal structure and function. The project centers on the MYST family of acetyltransferases that is implicated in normal growth and development and in conditions as diverse as leukemia, HIV-infection, and Alzheimer's disease. The proposed experiments build on results demonstrating that MYST proteins function in both transcriptional activation and silencing and have critical roles in DNA damage repair, cell cycle control, stationary phase survival, and chromosomal and subnuclear architecture. Previous studies focused on the three MYST genes, ESA1, SAS2 and SAS3. Planned studies will test the hypothesis that distinct roles for the MYST enzymes are specified through genomic targeting, mediated by interacting proteins and coordination with other chromatin modifying proteins. Transcriptional assays, affinity assays, mutational analysis, chromatin immunoprecipitation and microarray experiments will be used in these tests. Genetic and physical interactions that contribute to the distinct functions of SAS3 and ESA1 will be identified. Mechanisms of cell cycle control and DNA damage for SAS3 and ESA1 will be pursued to evaluate transcriptional vs. structural effects. These experiments will monitor checkpoint functions and will validate and extend microarray data. Cross-complementation experiments will be performed to address the specificity and extent of functional conservation of human MYST proteins. Preferred histone substrates have been identified for the yeast MYST enzymes, yet non-histone substrates are likely to be key to diverse MYST functions. Proteomic approaches will identify non-histone substrates, the significance of which will be evaluated through combined biochemical and genetic approaches.

描述（由申请人提供）：本研究的长期目标是了解染色质修饰调节基因表达和染色体结构与功能的机制。该项目以MYST乙酰转移酶家族为中心，该家族与正常生长和发育以及白血病，HIV感染和阿尔茨海默病等多种疾病有关。拟议的实验建立在结果表明，MYST蛋白的功能在转录激活和沉默，并在DNA损伤修复，细胞周期控制，稳定期生存，染色体和亚核结构中具有关键作用。以前的研究集中在三个MYST基因，ESA 1，SAS 2和SAS 3。计划中的研究将检验以下假设：MYST酶的不同作用是通过基因组靶向、相互作用蛋白质介导以及与其他染色质修饰蛋白的协调来指定的。在这些试验中将使用转录测定、亲和测定、突变分析、染色质免疫沉淀和微阵列实验。将确定有助于SAS3和ESA 1的不同功能的遗传和物理相互作用。将研究SAS 3和ESA 1的细胞周期控制和DNA损伤机制，以评估转录与结构效应。这些实验将监测检查点功能，并将验证和扩展微阵列数据。将进行交叉互补实验以解决人MYST蛋白的特异性和功能保守程度。已经确定了酵母MYST酶的优选组蛋白底物，但非组蛋白底物可能是多种MYST功能的关键。蛋白质组学方法将确定非组蛋白底物，其意义将通过生物化学和遗传学方法相结合进行评估。