Histone Modifying Enzymes and Transcription

组蛋白修饰酶和转录

• 批准号: 6989151
• 负责人: Rolf Sternglanz
• 金额: $ 29.76万
• 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 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6989151
• 关键词: Saccharomyces cerevisiae; acetylation; amidohydrolases; chromatin; cyclin dependent kinase; enzyme mechanism; fungal proteins; gene expression; gene mutation; genetic transcription; histones; mass spectrometry; methylation; methyltransferase; posttranslational modifications; temperature sensitive mutant

DESCRIPTION (provided by applicant): The long term goal of this research is to understand the role of histone modifications such as acetylation and methylation and their effect on chromosome function, using the widely-studied yeast, Saccharomyces cerevisiae, to study these processes. In particular this grant will focus on the effect of histone modifications on transcription. The grant has 3 aims. The first one (1) focuses on how the cyclin-dependent kinase Bur1 controls methylation of histone H3 lysine 36 by the histone methylase Set2. Set2 is known to be associated with RNA polymerase II during transcriptional elongation, and we think that Bur1 controls a crucial step in the elongation process, acting through Set2. The second aim is concerned with using mass spectroscopy on highly purified yeast histones H2A and H2B to identify modifications such as acetylation and methylation of lysine residues. Once specific modified residues have been identified, the effects of mutating them will be investigated and the enzymes responsible for the modifications will be determined. The third aim is to study Spt10, a protein known to be important for transcription of histone genes and perhaps other genes. Spt10 is thought to be an N-acetyltransferase based on its sequence, but its substrate may not be histones. A novel method involving peptide libraries will be used to identify the protein(s) that Spt10 acetylates. Since histones and their modifications are highly conserved from yeast to humans, as is the process of gene expression, these studies will lead to a greater understanding of fundamental functions of chromosomes in human cells.

描述(申请人提供)：这项研究的长期目标是了解组蛋白修饰的作用，如乙酰化和甲基化，以及它们对染色体功能的影响，使用被广泛研究的酵母，酿酒酵母，来研究这些过程。特别是，这项拨款将侧重于组蛋白修饰对转录的影响。这笔赠款有三个目标。第一个(1)集中在细胞周期蛋白依赖的激酶Bur1如何通过组蛋白甲基化酶Set2控制组蛋白H3赖氨酸36的甲基化。已知Set2在转录延伸过程中与RNA聚合酶II相关，我们认为Bur1通过Set2控制着延伸过程中的关键步骤。第二个目标是使用高纯度酵母组蛋白H_2A和H_2B的质谱学来鉴定修饰，例如赖氨酸残基的乙酰化和甲基化。一旦确定了特定的修饰残留物，将研究突变它们的影响，并确定负责修饰的酶。第三个目标是研究Spt10，这是一种已知对组蛋白基因转录很重要的蛋白质，或许还有其他基因。根据其序列，Spt10被认为是一种N-乙酰转移酶，但其底物可能不是组蛋白。一种涉及肽库的新方法将被用来鉴定Spt10乙酰化的蛋白质(S)。由于组蛋白及其修饰从酵母到人类都是高度保守的，就像基因表达的过程一样，这些研究将有助于更好地理解人类细胞中染色体的基本功能。