Role of POT1 in telomere length regulation

POT1 在端粒长度调节中的作用

• 批准号: 10618842
• 负责人: Sandy S Chang
• 金额: $ 33.5万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-09 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10618842
• 关键词: ATR checkpoint; Address; Adopted; Aging; Amino Acids; Binding; Binding Proteins; Biochemical; Biological Assay; Bone Marrow Transplantation; Bone marrow failure; Cell Aging; Cell Line; Cell Maintenance; Cell Proliferation; Cells; Chromosomal Rearrangement; Chromosome abnormality; Complex; Cytoprotection; DNA Binding; DNA Damage; DNA Double Strand Break; DNA biosynthesis; DNA replication fork; Data; Development; Disease; Dyskeratosis Congenita; Excision; Failure; Functional disorder; Future; Generations; Genes; Genetic; Genome; Genome Stability; Goals; Hematopoietic stem cells; Homeostasis; Homologous Gene; Human; In Vitro; Knockout Mice; Length; Malignant Neoplasms; Mediating; Metabolism; Molecular; Mus; Mutate; Mutation; Nature; Pathway interactions; Phenotype; Play; Premature aging syndrome; Property; Proteins; Public Health; Regulation; Repression; Role; SS DNA BP; Single-Stranded DNA; Structure; TP53 gene; Telomerase; Telomere Length Maintenance; Telomere Maintenance; Telomere-Binding Proteins; Testing; Therapeutic Studies; cancer initiation; genetic approach; in vivo; loss of function; mouse genome; mouse model; mutant; novel; premature; prevent; protein function; recruit; replication factor A; replication stress; response; single molecule; stem cell function; stem cells; telomere

Project Summary We will use state-of-the-art biochemical, cellular and genetic approaches to understand mechanistically how ssDNA binding proteins protect telomeres from activating a DNA damage response, regulate its length and replication. Using novel in vitro biochemical assays, we will examine whether the molecular distinctions between RPA and hPOT1 underlie their unique functions at telomeres. We will define the residues in hPOT1 that are required to repress telomere end protection and C-strand resection (Aim 1). We will determine mechanistically how POT1b promotes telomere length maintenance, and use hematopoietic stem cells conditionally depleted of endogenous POT1a/b to define the amino acid residues involved in telomere elongation necessary for the stem cell maintenance (Aim 2). We will explore how POT1a protects telomeres from replication stress. Finally, we used BioID to identify Claspin as a telomere interacting protein involved in DNA replication at telomeres devoid of POT1a (Aim 3). The proposed studies present a unique opportunity to address some of the most important questions in the telomere field.

项目概要 我们将使用最先进的生化、细胞和遗传学方法来了解 单链DNA结合蛋白如何保护端粒免于激活DNA损伤 响应，调节其长度和复制。使用新颖的体外生化测定，我们将 检查 RPA 和 hPOT1 之间的分子差异是否是其独特的基础 在端粒上发挥作用。我们将定义 hPOT1 中抑制端粒所需的残基 末端保护和 C 链切除（目标 1）。我们将机械地确定 POT1b 如何 促进端粒长度维持，并使用条件性耗竭的造血干细胞 内源性 POT1a/b 来定义参与端粒延长的氨基酸残基 干细胞维持所必需的（目标 2）。我们将探讨 POT1a 如何保护端粒 来自复制压力。最后，我们使用 BioID 鉴定 Claspin 为端粒相互作用蛋白 参与缺乏 POT1a 的端粒 DNA 复制（目标 3）。拟议的研究提出 这是解决端粒领域一些最重要问题的独特机会。