Isolation and Characterization of Yeast rDNA Chromatin

酵母 rDNA 染色质的分离和表征

• 批准号: 7155837
• 负责人: ERICA Y JACOBS
• 金额: $ 4.6万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7155837
• 关键词: Saccharomyces cerevisiae; binding sites; chimeric proteins; chromatin; electrospray ionization mass spectrometry; fungal genetics; genetic regulation; genetic transcription; green fluorescent proteins; histones; immunoprecipitation; matrix assisted laser desorption ionization; nucleic acid purification; nucleic acid repetitive sequence; postdoctoral investigator; posttranslational modifications; ribosomal DNA; transcription factor

DESCRIPTION (provided by applicant): How do genes turn on and off? If DNA sequence remains unchanged, changes in transcription are caused by differences in DNA-associated factors. So understanding chromatin composition changes within particular regions of DNA is vital to understanding gene expression. This proposal aims to better understand the role of chromatin dynamics in gene expression by examining how ribosomal DNA (rDNA) chromatin composition relates to rDNA transcription in the yeast Saccharomyces cerevisiae using mass spectrometry. This project has three specific aims. The first is to purify native rDNA chromatin from yeast cells. Second, the identity [and locations] of bound proteins and nature of histone modifications will be determined using mass spectrometry. Third, differences in chromatin composition and histone modifications between actively transcribed and silent rDNA will be determined. Future directions include extending the technology from yeast to mammalian cells. Chromatin composition at crucial loci could be followed through developmental transudations, environmental changes, or during oncogenesis. My ultimate goal is the complete characterization of an organism's "chromatome:" the identity, quantity, and location of all proteins associated with every DNA sequence and their modification states through development, aging and disease.

描述（由申请人提供）：基因如何打开和关闭？如果DNA序列保持不变，转录的变化是由DNA相关因子的差异引起的。因此，了解DNA特定区域内染色质组成的变化对了解基因表达至关重要。这项建议的目的是更好地了解染色质动力学在基因表达中的作用，通过研究如何核糖体DNA（rDNA）染色质组成与rDNA转录在酵母酿酒酵母中使用质谱。该项目有三个具体目标。第一个是从酵母细胞中纯化天然rDNA染色质。其次，结合蛋白的身份[和位置]和组蛋白修饰的性质将使用质谱法确定。第三，将确定活跃转录和沉默rDNA之间的染色质组成和组蛋白修饰的差异。未来的发展方向包括将该技术从酵母扩展到哺乳动物细胞。在关键位点的染色质组成可以通过发育渗漏，环境变化，或在肿瘤发生。我的最终目标是完整表征生物体的“染色体组”：与每个DNA序列相关的所有蛋白质的身份，数量和位置，以及它们在发育，衰老和疾病中的修饰状态。