Histone Modifications Associated w/Senescence in Arabidopsis

描述（由申请人提供）：该项目的长期目标是确定与拟南芥衰老相关的表观基因组。将收获非衰老和衰老叶组织，并使用染色质免疫沉淀和 Solexa/Illumina 高通量测序 (ChIP-SEQ) 来定义在全基因组范围内与修饰组蛋白结合的 DNA 区域。基因表达也将通过 ChIP-SEQ 使用识别 RNA 聚合酶 II 的抗体在全基因组范围内进行评估。待分析的组蛋白修饰包括甲基化、乙酰化和泛素化。组蛋白脱乙酰酶的过度表达会延缓人类细胞的衰老，最近发现拟南芥中的延迟衰老突变体 ore7-1D 是由 AT-hook 染色质修饰蛋白的过度表达引起的。 ore7-1D突变体的表观基因组将在Specific Aim 4中进行评估，这可能会导致深入了解植物和人类之间共享的细胞衰老的表观遗传调控。该试点提案的另一个目标是通过与附近 R01 大学的一位成功的表观基因组研究员建立持续的指导关系，提高 PI 的研究竞争力。

DESCRIPTION (provided by applicant): The long-term objective of this project is to define the epigenome associated with senescence in Arabidopsis thaliana. Non-senescent and senescent leaf tissue will be harvested, and chromatin immunoprecipitation followed by Solexa/Illumina high throughput sequencing (ChIP-SEQ) will be used to define DNA regions that bind to modified histones on a genome-wide scale. Gene expression will also be evaluated on a genome-wide scale by ChIP-SEQ using an antibody that recognizes RNA Polymerase II. Histone modifications to be analyzed include methylations, acetylations and ubiquitinations. Cellular aging in humans is delayed by overexpression of histone deacetylases, and a delayed senescence mutant in Arabidopsis, ore7-1D, was recently shown to result from overexpression of an AT-hook chromatin modifying protein. The epigenome of the ore7-1D mutant will be assessed in Specific Aim 4, which may lead to insights into epigenetic regulation of cellular aging that are shared between plants and humans. Another goal of this pilot proposal is to increase the research competitiveness of the PI via an ongoing mentoring relationship with a successful epigenome researcher at a nearby R01 university.