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

ATP依赖的染色质重塑复合物，如进化上保守的INO 80复合物， 对维持基因组完整性至关重要。然而，不同的INO 80染色质修饰 正确的DNA损伤反应所必需的活性仍有待阐明。我们的长期目标 描述染色质修饰复合物在DNA损伤反应途径中的参与， 以及它们通过维持基因组完整性在预防疾病进展中的作用。的 本提案的总体目标是实现我们长期目标的第一步， 酵母模型系统，以确定INO 80染色质重塑复合物的具体参与， DNA损伤反应途径的不同功能。我们的中心假设是，特定的INO 80 亚基在不同的基因组维持途径中发挥关键作用， 复合物的染色质修饰活性。这些目标一旦实现， 为研究进化保守的染色质重塑复合物在 由DNA损伤反应途径缺陷引起的疾病，如癌症。以强为导 初步数据，这一假设将通过一个综合的实验方法，利用 蛋白质组学、高通量遗传筛选、全基因组核小体作图和生物化学测定。 我们提出通过追求以下具体目标来检验我们的中心假设并实现本申请的目标：1）表征在DNA损伤反应中起作用并调节DNA损伤特异性染色质底物的INO 80亚基：2）描绘DNA损伤相关亚基对INO 80复合物活性的体外染色质修饰贡献; 3）确定INO 80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来解决INO 80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节的理解，染色质调节的功能是保持DNA的完整性。

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.