Defining the cellular functions of the conserved transcription complex Spt6/Iws1 in the control of gene expression

定义保守转录复合体 Spt6/Iws1 在控制基因表达中的细胞功能

• 批准号: 9284260
• 负责人: Olga V. Viktorovskaya
• 金额: $ 5.92万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-01 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9284260
• 关键词: Affect; Alleles; Animal Model; Antineoplastic Agents; Biochemical; Cell Differentiation process; Cell physiology; Cells; ChIP-seq; Chromatin; Chromatin Structure; Complex; Defect; Development; Disease; Enzymes; Eukaryota; Event; Failure; Future; Gene Expression; Genes; Genetic; Genetic Screening; Genetic Transcription; Goals; Growth; Heterodimerization; Histones; Human; Human Biology; Human Characteristics; Lead; Learning; Light; Link; Malignant Neoplasms; Measurement; Messenger RNA; Mutation; Nature; Nucleosomes; Oncogene Deregulation; Pathologic; Pathway Analysis; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Play; Process; Proteins; RNA Polymerase II; RNA Processing; Recruitment Activity; Regulation; Research Project Grants; Role; Saccharomycetales; Sequence Homology; Signal Transduction; Site-Directed Mutagenesis; Suppressor Mutations; Techniques; Testing; Therapeutic Intervention; Transcription Elongation; Transcriptional Regulation; Yeasts; base; cell growth; drug discovery; environmental change; genome-wide; histone modification; human disease; in vivo; mutant; neoplastic cell; novel; prevent; protein protein interaction; public health relevance; response; success; targeted treatment; therapeutic target; transcription factor; transcriptome sequencing; tumor; tumorigenesis

 DESCRIPTION (provided by applicant): Transcriptional regulation is an essential aspect of cell growth and differentiation, as well as critical in the response of cells to environmental changes. In humans, defects in transcription often lead to disease. Therefore, the elucidation of transcriptional mechanisms is important for understanding human biology. This proposal focuses on two proteins, Spt6 and Iws1, that are conserved from yeast to humans, that are essential for viability, and that have been implicated in cancer. Spt6 and Iws1 function as global transcriptional regulators and control gene expression at multiple levels. Despite intensive studies, the exact mechanisms by which Spt6/Iws1 regulates transcription is poorly understood. To investigate the roles that Spt6 and Iws1 play in gene expression the following specific aims are proposed: Aim 1. Elucidate the in vivo roles of the conserved motifs of the Iws1 protein. Using site- directed mutagenesis of the conserved residues, isolation of conditionally lethal mutants, and a protein depletion approach, the functions of Iws1 will be analyzed. The proposed mutant analysis will reveal the functions of Iws1 in transcription, mRNA processing, protein-protein interactions, and recruitment of critical factors to chromatin genome-wide. Aim 2. Isolate and characterize suppressors of spn1Δ and the spt6Δ null alleles. A genetic screen for suppressors of iws1Δ and spt6Δ inviability will be performed to discover novel cellular activitie and pathways that can compensate for loss of Spt6 and Iws1. Detailed characterization of the identified suppressors, including several mutations already identified in the pilot screen, will shed light on the mechanisms by which these mutants rescue absence of Iws1 and will expand our understanding on the roles of Spt6 and Iws1 in gene expression. Altogether, the proposed studies will further our current understanding of the essential functions of Spt6 and Iws1 in transcriptional regulation. Our long-term goal is to understand the cascade of events and the contribution of each activity in gene expression control. Relevance: Deregulation of gene expression is tightly linked to tumorigenesis. Given the conserved nature of the studied factors and their roles in development and oncogenesis, our studies using yeast as model organisms will be broadly applicable to other eukaryotes. This research project has a long-term implication in anticancer drug discovery.

 描述（由申请人提供）：转录调控是细胞生长和分化的一个重要方面，也是细胞对环境变化反应的关键。在人类中，转录缺陷往往导致疾病。因此，阐明转录机制对于理解人类生物学是重要的。该提案的重点是两种蛋白质，Spt 6和Iws 1，这两种蛋白质从酵母到人类都是保守的，对生存能力至关重要，并且与癌症有关。Spt 6和Iws 1作为全局转录调节因子，在多个水平上控制基因表达。尽管深入研究，Spt 6/Iws 1调节转录的确切机制知之甚少。为了研究Spt 6和Iws 1在基因表达中发挥的作用，提出了以下具体目标：目标1。阐明Iws 1蛋白保守基序在体内的作用。使用保守残基的定点突变、条件致死突变体的分离和蛋白质耗尽方法，将分析Iws 1的功能。拟议的突变体分析将揭示Iws 1在转录，mRNA加工，蛋白质-蛋白质相互作用，和招聘的关键因素染色质全基因组的功能。目标2.分离和表征sture 1 Δ和spt 6 Δ无效等位基因的抑制子。将对iws 1 Δ和spt 6 Δ失活的抑制因子进行遗传筛选，以发现新的细胞活性和途径，从而补偿Spt 6和Iws 1的缺失。详细表征所确定的抑制因子，包括几个突变已经确定的试点屏幕，将揭示这些突变体救援缺乏Iws 1的机制，并将扩大我们的理解Spt 6和Iws 1在基因表达中的作用。总之，拟议的研究将进一步我们目前的理解Spt 6和Iws 1的转录调控的基本功能。我们的长期目标是了解级联事件和基因表达控制中每个活动的贡献。相关性：基因表达失调与肿瘤发生密切相关。鉴于所研究的因子的保守性及其在发育和肿瘤发生中的作用，我们使用酵母作为模式生物的研究将广泛适用于其他真核生物。该研究项目在抗癌药物发现方面具有长期意义。