Control of Programmed Replication fork Arrest by Chromosome Kissing

通过染色体亲吻控制程序化复制叉停滞

• 批准号: 8638786
• 负责人: DEEPAK BASTIA
• 金额: $ 40.2万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8638786
• 关键词: 3-Dimensional; Accounting; Address; Alleles; Binding Sites; Biological; CDC45L gene; Cells; Chromatin Loop; Chromosomal translocation; Chromosome Pairing; Chromosomes; Complex; DNA; DNA biosynthesis; DNA-Protein Interaction; Development; Drosophila genus; Epigenetic Process; Fission Yeast; Gene Expression; Gene Silencing; Genes; Genetic Recombination; Genetic Screening; Genetic Transcription; Goals; Histone Deacetylase; In Vitro; Interphase; Investigation; Laboratories; MCM2 gene; Malignant Neoplasms; Mammalian Cell; Mediating; Mitosis; Modeling; Molecular Conformation; Movement; Nucleosomes; Paper; Physiological; Prevention; Process; Proteins; Publishing; Recombinant DNA; Reporting; Role; Schizosaccharomyces pombe Proteins; Site; Techniques; Testing; Time; Work; X Inactivation; cancer cell; chromatin remodeling; deep sequencing; genome wide association study; genome-wide; helicase; human disease; mutant; novel; prevent; programs; promoter; reconstitution; research study; two-dimensional

DESCRIPTION (provided by applicant): Major DNA transactions such as transcription, development, X-chromosome inactivation and chromosomal translocations in cancer cells were reported to be controlled by chromosome to chromosome contacts called "chromosome kissing" mediated by a myb-like terminator protein called Reb1. We have recently shown in a paper to be published in Cell that chromosome kissing in fission yeast also controls replication fork movement by controlling sequence-specific, physiologically programmed fork arrest. This finding raises some major questions: (i) which proteins are involved in controlling chromosome kissing and what are their mechanisms of action?; (ii) what is the mechanism of programmed fork arrest? In the context of addressing the first question, we have discovered that two chromatin remodeling proteins of fission yeast modulate programmed fork arrest. The implication is that these remodelers do this by modulating chromosome kissing and nucleosome disposition about the replication termini. One of the goals of this proposal is to discover in a genome-wide search the proteins that modulate chromosome kissing and try to understand their mechanism of action. Another important goal of this proposal is to uncover the mechanism of action of programmed fork arrest by testing the hypothesis that this happens by unidirectional arrest of the MCM2-7 helicase. Programmed fork arrest controls other chromosome transactions such as recombination, gene silencing and transcriptional passage and is at the interphase of replication and other processes. Finally, experiments are proposed to localize genome-wide Reb1-dependent replication termini. The objectives of this goal is to test two hypotheses: (i) naturally occurring, noncanonical weak sites are rendered functional by chromosome kissing and/or DNA looping -dependent interaction with strong canonical sites and that is one function of these long range protein- DNA interactions; (ii) a function of looping-dependent (or independent) fork arrest is to prevent interference between replication and transcription.

描述（由申请人提供）：据报道，癌细胞中的主要 DNA 事务，如转录、发育、X 染色体失活和染色体易位，是由称为“染色体接吻”的染色体间接触控制的，该接触由称为 myb 的终止子蛋白介导。雷布1。我们最近在《细胞》杂志上发表的一篇论文中表明，裂殖酵母中的染色体亲吻还通过控制序列特异性、生理程序化的复制叉停滞来控制复制叉的运动。这一发现提出了一些主要问题：（i）哪些蛋白质参与控制染色体亲吻以及它们的作用机制是什么？ (ii) 程序化叉子制动的机制是什么？在解决第一个问题的背景下，我们发现裂殖酵母的两种染色质重塑蛋白调节程序性分叉停滞。这意味着这些重塑者通过调节复制末端的染色体亲吻和核小体配置来实现这一点。该提案的目标之一是在全基因组搜索中发现调节染色体亲吻的蛋白质，并尝试了解它们的作用机制。该提案的另一个重要目标是通过测试通过 MCM2-7 解旋酶的单向抑制发生的假设来揭示程序化叉抑制的作用机制。程序化的分叉停滞控制其他染色体事务，例如重组、基因沉默和转录通道，并且处于复制和其他过程的中间期。最后，提出了实验来定位全基因组 Reb1 依赖的复制末端。这一目标的目的是检验两个假设：(i) 天然存在的非规范弱位点通过染色体亲吻和/或 DNA 循环依赖性与强规范位点的相互作用而发挥功能，这是这些长程蛋白质-DNA 的功能之一。互动； (ii)环依赖性（或独立）的叉子停滞的功能是防止复制和转录之间的干扰。