RSF: properties and dependence on histone modifications

RSF：组蛋白修饰的特性和依赖性

• 批准号: 7232553
• 负责人: Dawn M Schmidt
• 金额: $ 0.7万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-14 至 2007-07-13
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7232553
• 关键词: adenosine triphosphate; carcinogenesis; chromatin; enzyme complex; genetic disorder; histones; hydrolysis; molecular assembly /self assembly; nucleoproteins; nucleosomes; phosphatidylinositols; postdoctoral investigator; protein binding; protein protein interaction; protein structure function; site directed mutagenesis

DESCRIPTION (provided by applicant): Defects in chromatin remodeling factors and histone modifying enzymes are associated with tumorigenesis and other genetic disorders. Therefore it is critical to understand these activities and their influence on one another. The human chromatin remodeling complex RSF (remodeling and spacing factor) is composed of two subunits, the novel protein Rsf-1 and the ISWI-type ATPase hSNF2H. RSF does not require exogenous histone chaperones for nucleosome assembly and spacing. The influence of specific histone modifications on the function of the individual subunits has not been determined; this will be accomplished through the use of native chemical ligation to synthesize site-specifically modified histones, as well as histones incorporating biophysical probes. In particular, the influence of these modifications on nucleosome assembly versus spacing will be assessed. The timing of the coupling of ATP hydrolysis to chromatin assembly events will also be examined. The function of Rsf-1 will be probed through mutagenesis, initially of the PHD zinc finger domain common to many chromatin-associated proteins. Rsf-1 and its PHD domain will be probed for their ability to bind phosphoinositols, and if so, for their influence on RSF function.

描述（由申请人提供）： 染色质重塑因子和组蛋白修饰酶的缺陷与肿瘤发生和其他遗传性疾病有关。因此，了解这些活动及其相互影响至关重要。人类染色质重塑复合物 RSF（重塑和间隔因子）由两个亚基组成，即新型蛋白 Rsf-1 和 ISWI 型 ATP 酶 hSNF2H。 RSF 不需要外源组蛋白伴侣来进行核小体组装和间隔。特定组蛋白修饰对各个亚基功能的影响尚未确定；这将通过使用天然化学连接来合成位点特异性修饰的组蛋白以及结合生物物理探针的组蛋白来实现。特别是，将评估这些修饰对核小体组装与间距的影响。还将检查 ATP 水解与染色质组装事件耦合的时间。 Rsf-1 的功能将通过诱变来探测，首先是许多染色质相关蛋白共有的 PHD 锌指结构域。将探测 Rsf-1 及其 PHD 结构域结合磷酸肌醇的能力，如果是这样，则探测它们对 RSF 功能的影响。