Histone Modifications Associated w/Senescence in Arabidopsis

描述（由申请人提供）：本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，然后使用Solexa/Illumina高通量测序（ChIP-SEQ）来定义在全基因组范围内结合修饰的组蛋白的DNA区域。还将使用识别RNA聚合酶II的抗体通过ChIP-SEQ在全基因组范围内评价基因表达。待分析的组蛋白修饰包括甲基化、乙酰化和泛素化。人类的细胞衰老可通过组蛋白去乙酰化酶的过度表达而延缓，最近发现拟南芥中的延迟衰老突变体ore 7 -1D是由AT-钩染色质修饰蛋白的过度表达引起的。ore 7 -1D突变体的表观基因组将在特定目标4中进行评估，这可能会导致对植物和人类之间共享的细胞衰老的表观遗传调控的深入了解。该试点提案的另一个目标是通过与附近R 01大学成功的表观基因组研究人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.