Epigenetic and Metabolic Regulation of Gene Silencing in Saccharomyces

Mira奖的申请旨在巩固PI实验室的研究，资金来源为 GM31105对控制位置效应的基因的遗传分析，目前已进入第37个年头，并通过 GM120374新陈代谢和表观遗传学，目前已进入第四个年头，纳入一个计划。这项工作 在这些赠款的支持下，产生了一系列关于以下方面的基本发现 异色基因沉默的机制、表观遗传及其对 新陈代谢。最近，这些努力还包括开发变革性新的 基因技术，并成功启动了一项全面调查 关于表观遗传过程的代谢，最初的重点是那些改变新陈代谢的突变 是人类癌症的驱动因素。GM31105的长期遗产使人们能够发现SIR基因是如何 控制异染色质的形成，沉默分子在控制基因表达中的作用，以及 转录状态的表观遗传。在这个过程中有许多附带的发现 研究包括(1)定义起源识别复合体的第一个突变，及其可分离的 在DNA复制和调控转录中的作用；(2)确定 异染色质；(3)揭示异染色质动力学的单细胞分析；以及(4)发现 不同寻常的不育突变导致了a因子和RAS的蛋白质预烯基化的发现。最近， GM120374支持的工作导致发现克雷布斯循环各组成部分的突变 作为癌症驱动因素的突变对组蛋白去甲基酶和基因有意想不到的影响 沉默，并发现了与异染色质稳定性有关的新的代谢联系。这些赠款加在一起，有 产生了一种变革性的技术，用于单个核小体的位点特异性标记 允许解决关于染色质的最长的悬而未决的问题之一，表明 单个核小体通过多个DNA复制周期保持其基因组地址。这个 来自这些拨款的最新数据迫使人们从根本上重新考虑核小体是否是 表观遗传状态记忆的载体。 拟议的研究计划将回答表观遗传学中的多个长期存在的问题 例如确定允许表观遗传的存储器组件驻留在哪里 转录状态和分解记忆的机制。此外，拟议的研究 将解决基因沉默从其成核部位横向传播的机制。这个 创造新的特定细胞类型的基因表达状态的细胞周期要求将是 已确认身份。最后，新发现的代谢对表观遗传学影响的机制 沉默将被确定。

This application for a MIRA award seeks to consolidate the research in the PI's lab, funded by GM31105 Genetic Analysis of Genes Controlling a Position Effect, currently in its 37th year, and by GM120374 Metabolism and Epigenetics, currently in its fourth year, into one program. The work supported by these grants has produced a continuous series of fundamental discoveries about mechanisms of heterochromatic gene silencing, its epigenetic inheritance, and its sensitivity to metabolism. Most recently, these efforts have also included the development of transformative new genetic technologies, and the successful launch of a comprehensive investigation of the impact of metabolism on epigenetic processes, with an initial focus on those metabolism-altering mutations that are drivers of human cancers. The long legacy of GM31105 enabled the discovery of how the Sir genes control heterochromatin formation, the role of silencers in controlling gene expression, and the epigenetic inheritance of transcriptional states. Many ancillary discoveries made in the course of these investigations include (1) the first mutations defining the Origin Recognition Complex, and its separable roles in DNA replication and regulating transcription; (2) defining the molecular topography of heterochromatin; (3) single-cell assays revealing heterochromatin dynamics; and (4) discovery of unusual sterile mutations that led to the discovery of protein prenylation of a-factor and Ras. Recently, the work supported by GM120374 led to the discovery that mutations in components of the Krebs cycle that function as cancer driver mutations have unexpected impacts on histone demethylases and gene silencing, and identified new metabolic links to heterochromatin stability. Together, these grants have produced a transformative technique for the locus-specific labeling of individual nucleosomes that allowed resolution of one of the longest unsolved questions regarding chromatin, showing that individual nucleosome retain their genomic addresses through multiple DNA replication cycles. The newest data from these grants force a fundamental reconsideration of whether nucleosomes are the carriers of the memory of epigenetic states. The proposed research program will answer multiple long-standing questions in epigenetics such as determining where the memory component resides that allows epigenetic inheritance of transcriptional states and resolving the mechanism of that memory. In addition, the proposed research will resolve the mechanism by which gene silencing spreads laterally from its sites of nucleation. The cell-cycle requirements for the creation of new cell-type-specific states of gene expression will be identified. Finally, the mechanism by which the newly identified metabolic impacts on epigenetic silencing will be determined.