MOLECULAR INTERACTIONS AND SUBSTRATES OF MAMMALIAN SIRT6 LONGEVITY REGULATOR

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 Sir 2（Silent Information Regulator-2）蛋白在多种模式生物中的衰老和寿命调节中起作用。 初步研究表明，一种哺乳动物Sir 2蛋白，SIRT 6，抑制基因组不稳定性，并减弱了几种年龄相关的病理学的发病。 然而，SIRT 6功能的分子机制尚不清楚。 因此，SIRT 6在人类细胞中发挥作用的基本分子机制的系统表征应该有助于阐明影响人类健康和衰老的基本生物学过程。 将进行一系列生物化学、细胞和全球蛋白质组学和基因组学分析，以确定SIRT 6在人类细胞中的分子功能。 我们建议： 1. 阐明SIRT 6在染色质调控中发挥作用的分子机制。 组蛋白在染色质上的翻译后修饰在基因表达和DNA损伤反应中起着重要作用。 初步结果表明，SIRT 6与染色质上的组蛋白紧密相关，并可能催化组蛋白的修饰。 我们将定义SIRT 6在染色质上的酶活性以及改变的SIRT 6水平对染色质结构和功能状态的影响。 2. 阐明SIRT 6调控的新分子通路。 将采取蛋白质组学和生物化学方法来分离和鉴定SIRT 6底物、蛋白质结合伴侣和大分子复合物。 此外，将采用全基因组方法来鉴定SIRT 6调节的基因和染色质上的SIRT 6结合位点。 与UCSF质谱设施合作的质谱分析将对这些目标至关重要，在鉴定SIRT 6催化的组蛋白和其他蛋白质的翻译后修饰，以及SIRT 6相互作用因子和大分子复合物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Sir2 (Silent Information Regulator-2) proteins function in aging and lifespan regulation in multiple model organisms. Preliminary work indicates that one mammalian Sir2 protein, SIRT6, suppresses genomic instability and attenuates the onset of several age-associated pathologies. However, the molecular mechanisms of SIRT6 function are not known. Thus, a systematic characterization of the basic molecular mechanisms through which SIRT6 functions in human cells should be instrumental for elucidating fundamental biological processes that impact on human health and aging. A series of biochemical, cellular, and global proteomic and genomic analyses will be carried out to define the molecular functions of SIRT6 in human cells. We propose: 1. To elucidate the molecular mechanisms by which SIRT6 functions in chromatin regulation. Post-translational modifications of histones at chromatin play important roles in gene expression programs and DNA damage responses. Preliminary results indicate that SIRT6 is tightly associated with, and may catalyze modifications of, histones at chromatin. We will define the enzymatic activity of SIRT6 at chromatin and the effects of altered SIRT6 levels on chromatin structure and functional states. 2. To elucidate novel SIRT6 regulated molecular pathways. Proteomic and biochemical approaches will be taken to isolate and identify SIRT6 substrates, protein binding partners, and macromolecular complexes. In addition, genome wide approaches will be employed to identify SIRT6-regulated genes and SIRT6 binding sites at chromatin. Mass Spectrometry analysis in collaboration with the UCSF Mass Spectrometry Facility will be essential for these aims, in the identification of post-translational modifications on histones and other proteins catalyzed by SIRT6, and of SIRT6 interacting factors and macromolecular complexes.