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: 8206077
• 负责人: DEEPAK BASTIA
• 金额: $ 41.7万
• 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 2015-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8206077
• 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; 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. PUBLIC HEALTH RELEVANCE: The existing models of replication control are two dimensional consisting of proteins that interact with sequences of a single chromosome at a time to initiate replication, promote fork progression etc. Our recent work shows that replication control is 3 dimensional. This application proposes to investigate further this mechanism that is potentially relevant to human diseases such as cancer.

描述（由申请人提供）：据报道，癌细胞中的主要DNA交易，例如转录，发育，X染色体失活和染色体易位，由染色体与称为“染色体接吻”的染色体控制，称为“染色体接吻”，该接触被myb样末端末端蛋白称为Reb1。最近，我们在旨在在细胞中发表的论文中表明，裂变酵母中的染色体接吻还通过控制序列特异性，生理编程的叉式停滞来控制复制叉运动。这一发现提出了一些主要问题：（i）哪些蛋白质涉及控制染色体接吻？它们的作用机理是什么？ （ii）编程叉逮捕的机制是什么？在解决第一个问题的背景下，我们发现裂变酵母调节编程的叉式叉子骤停的两种染色质重塑蛋白。这意味着这些重塑器是通过调节复制末端的染色体接吻和核小体处置来做到这一点的。该提案的目标之一是在全基因组搜索中发现调节染色体接吻的蛋白质并试图理解其作用机理。该提案的另一个重要目标是通过检验以下假设，即通过单向逮捕MCM2-7解旋酶来揭示编程叉骤停的作用机理。编程的叉式逮捕控制其他染色体交易，例如重组，基因沉默和转录通过，并且处于复制和其他过程的相间。最后，提出了实验来定位全基因组REB1依赖性复制末端。该目标的目的是检验两个假设：（i）自然发生的，非规范的弱位点通过染色体接吻和/或DNA循环依赖性的相互作用与强量典型位点具有功能，这是这些长范围蛋白质 - DNA相互作用的一个功能； （ii）依赖循环依赖（或独立的）叉的函数是防止复制和转录之间的干扰。 公共卫生相关性：现有的复制控制模型是由蛋白质组成的二维模型，该蛋白质一次与单个染色体的序列相互作用，以启动复制，促进叉进程等。我们最近的工作表明，复制控制是3维控制的。该应用建议进一步研究这种机制，该机制可能与癌症等人类疾病有关。