Chromatin-modifying functions of INO80 in the preservation of genomic integrity

依赖于ATP的染色质重塑复合体，如进化保守的INO80复合体，是 对于维持基因组的完整性是必不可少的。然而，不同的INO80染色质修饰 正确的DNA损伤反应所必需的活动仍有待阐明。我们的长期目标 是描绘染色质修饰复合体在DNA损伤反应通路中的参与，如 以及它们在通过维持基因组完整性防止疾病进展方面的作用。这个 这项建议的总体目标，是迈向我们长远目标的第一步，利用 酵母模型系统确定INO80染色质重塑复合体在 不同功能的DNA损伤反应通路。我们的中心假设是特定的INO80 亚基在不同的基因组维持途径中起关键作用，它通过调节 该络合物的染色质修饰活性。一旦实现了这些目标，结果将是 为研究进化保守的染色质重塑复合体在 由DNA损伤反应通路缺陷引起的疾病，如癌症。以Strong为指导 初步数据，这一假设将通过综合实验方法进行验证，该方法利用 蛋白质组学、高通量遗传筛选、全基因组核小体图谱和生化分析。 我们建议验证我们的中心假设，并通过追求以下特定目标来实现这一应用的目标：1)表征在DNA损伤反应中起作用的INO80亚基并调节DNA损伤特异性染色质底物；2)描述DNA损伤相关亚基对活性INO80复合体的染色质修饰作用；3)在体内确定INO80亚基在特定DNA损伤反应途径中的染色质修饰功能。这些分析与公众健康相关，因为它们将提供一个平台，以解决INO80在疾病发展中的潜在作用。最终，这些研究将极大地拓宽我们对染色质调制的理解，染色质调制具有保护DNA完整性的功能。

ATP-dependent chromatin-remodeling complexes, such as the evolutionarily-conserved INO80 complex, are essential for the maintenance of genomic integrity. However, the diverse INO80 chromatin-modifying activities that are necessary for proper DNA damage responses remain to be elucidated. Our long-term goal is to delineate the involvement of chromatin-modifying complexes in DNA damage response pathways, as well as their role in preventing disease progression through the maintenance of genomic integrity. The overall objective of this proposal, which is an initial step to the achievement of our long-term goal, utilizes the yeast model system to determine the specific involvement ofthe INO80 chromatin-remodeling complex in different functions of DNA damage response pathways. Our central hypothesis is that specific INO80 subunits contribute to critical functions in diverse genome maintenance pathways by modulating the chromatin-modifying activities of the complex. Once these objectives are accomplished, the results will provide the foundation to investigate the role of evolutionarily-conserved chromatin-remodeling complexes in disorders caused by defects in DNA damage response pathways, such as cancer. Guided by strong preliminary data, this hypothesis will be tested through an integrated experimental approach that utilizes proteomics, high-throughput genetic screens, genome-wide nucleosome mapping, and biochemical assays. We propose to test our central hypothesis and accomplish the objectives ofthis application by pursuing the following specific aims: 1) Characterize INO80 subunits that function in DNA damage responses and modulate DNA damage-specific chromatin substrates; 2) Delineate the in vitro chromatin-modifying contributions of DNA damage-related subunits to the activity INO80 complex; 3) Determine the in vivo chromatin-modifying function of INO80 subunits in specific DNA damage response pathways. These analyses are relevant to public heaith because they will provide a platform to address the potential role of INO80 in the development of disease. Ultimately, these studies will dramatically broaden our understanding of chromatin modulation that functions to preserve DNA integrity.