Role of Heterochromatin protein 1 Beta in Genome Maintenance and Oncogenesis

异染色质蛋白 1 Beta 在基因组维护和肿瘤发生中的作用

• 批准号: 9309045
• 负责人: Tej K Pandita
• 金额: $ 30.31万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-16 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9309045
• 关键词: Acetylation; Appearance; Applications Grants; Binding; Biochemical; C-terminal; Cancer Patient; Cell Survival; Cell physiology; Cells; Cessation of life; ChIP-on-chip; Chromatin; Chromatin Structure; Chromosomes, Human, Pair 1; Clinical; Co-Immunoprecipitations; Complement; Complex; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA lesion; DNA-PKcs; Data; Development; Double Strand Break Repair; Embryo; Euchromatin; Eukaryota; Fibroblasts; Frequencies; Genes; Genetic Transcription; Genome; Genomic Instability; Genomics; Goals; Heterochromatin; Histone H3; Histone H4; Histones; Human; Individual; Investigation; Ionizing radiation; Kinetics; Knockout Mice; Lasers; Link; Lysine; Maintenance; Mammalian Cell; Mammals; Mammary Gland Parenchyma; Mediating; Modification; Mus; N-terminal; NBS1 gene; Neoplastic Cell Transformation; Normal Cell; Normal tissue morphology; Oncogenic; Pathway interactions; Phenotype; Play; Post-Translational Protein Processing; Predisposition; Protein Isoforms; Proteins; Radiation Induced DNA Damage; Radiation Tolerance; Radiation exposure; Radiation therapy; Recruitment Activity; Reporting; Research; Research Personnel; Role; Site; Tail; Testing; Time; Tissues; Tumor Tissue; ataxia telangiectasia mutated protein; base; cell injury; cell killing; chromatin protein; expression vector; fluorescence imaging; heterochromatin-specific nonhistone chromosomal protein HP-1; histone modification; improved; in vivo; insight; novel; novel strategies; overexpression; p53-binding protein 1; prevent; public health relevance; radiation response; repaired; response; tumor; tumorigenesis; zinc finger nuclease

DESCRIPTION (provided by applicant): The DNA damage response (DDR) mediates DNA double strand break (DSB) repair and protects cells from damage induced transformation or death. Cellular DNA is organized into protein DNA complexes (chromatin) in order to control DNA access by proteins and regulated DNA dependent functions. In eukaryotes, there are two major types of chromatin: heterochromatin (gene poor) and euchromatin (gene rich) that are distinguished by specific histone tail modifications and differences in nonhistone chromatin protein constituents. Among nonhistone chromatin proteins, heterochromatin protein 1 (HP1) is the best- studied example. In mammals, there are three HP1 isoforms (HP1a, HP1b and HP1g) all structurally characterized by two conserved domains separated by a hinge region: an N-terminal chromodomain (CD) and a C-terminal chromoshadow domain (CSD). We previously demonstrated that overexpression of HP1a or HP1b in human cells increases genomic instability and sensitivity to ionizing radiation (IR)-induced cell killing. Moreover, depletion of Cbx1 (mouse HP1b) in mouse cells increased genomic instability, spontaneous ATM (ataxia-telangiectasia mutated) autophosphorylation, reduced the frequency of IR-induced g-H2AX foci formation, increased IR-induced cell killing and oncogenic transformation. Recent studies by other investigators support our results indicating HP1b has both negative as well as positive effects on DNA DSB repair and suggest that the precise level of functional HP1b is a critical determinant to IR sensitivity. Since most mechanistic details as to how HP1 b interacts with repair associated proteins to modulate DNA DSB repair are unexplored, we will determine how different domains interact with chromatin/repair protein components to regulate DNA DSB repair. We hypothesize that the negative effect of HP1b is mediated through CD domain binding to H3K9me, since deletion of this domain can improve cell survival and the positive effect could be due to HP1b CSD interactions with acetylated histone H4K16 (H4K16ac) a unique histone modification that prevents higher order chromatin packing, which can impede protein access to DNA and with proteins involved in the DDR. Defective DNA damage repair is linked with oncogenic transformation and tumorigenesis, therefore, we will determine the impact of decreased Cbx1 on tumor development in (i) Cbx1+/- mice in the presence and absence of Atm and (ii) Cbx1 conditional knockout mice. These studies will define the mechanism by which HP1b regulates the cellular IR response and tumorigenesis. Our hypothesis-that the non-histone modifying factor HP1b regulates chromatin structure and, through interactions with DDR components, contributes to oncogenesis-is a novel idea requiring in depth studies. Further understanding about the mechanistic basis for biochemical differences between normal and tumor tissue chromatin structure will facilitate the development of new strategies for modifying IR response and improving clinical radiation therapy.

描述（由申请人提供）：DNA损伤响应（DDR）介导DNA双链断裂（DSB）修复并保护细胞免受损伤诱导的转化或死亡。细胞DNA被组织成蛋白DNA复合物（染色质），以控制蛋白质和调节DNA依赖性功能的DNA访问。在真核生物中，有两种主要类型的染色质：异染色质（基因较差）和周染色质（富含基因），它们通过特异性组蛋白尾巴修饰和非组蛋白染色质蛋白蛋白成分的差异进行区分。在非组蛋白染色质蛋白中，杂染色质蛋白1（HP1）是最佳研究的例子。在哺乳动物中，有三种HP1同工型（HP1A，HP1B和HP1G）在结构上以两个由铰链区域分离的保守域进行特征：N末端染色体域（CD）和一个C-末端Chromoshadow域（CSD）。我们先前证明了人类细胞中HP1A或HP1B的过表达会增加基因组不稳定性和对电离辐射（IR）诱导的细胞杀伤的敏感性。而且，耗尽 小鼠细胞中的CBX1（小鼠HP1B）增加了基因组不稳定性，自发性ATM（共济失调 - 凝性突变）自磷酸化，降低了IR诱导的G-H2AX焦点形成的频率，增加了IR诱导的细胞杀死和致癌转化。其他研究者的最新研究支持我们的结果表明HP1B对DNA DSB修复具有负和积极影响，并表明功能性HP1B的精确水平是IR敏感性的关键决定因素。由于关于HP1 B如何与修复相关蛋白相互作用以调节DNA DSB修复的大多数机械细节，因此我们将确定不同域如何与染色质/修复蛋白成分相互作用以调节DNA DSB修复。我们假设HP1B的负面影响是通过CD结构域与H3K9ME结合的，因为该结构域的缺失可以提高细胞存活，并且阳性效应可能是由于HP1B CSD与乙酰化组蛋白H4K16（H4K16AC）的相互作用，可预测的一种独特的组蛋白修饰。高阶染色质填料，这会阻碍蛋白质进入DNA，并且与DDR有关的蛋白质涉及蛋白质。 DNA损伤修复有缺陷与致癌性转化和肿瘤发生有关，因此，我们将确定CBX1减少对（I）CBX1 +/-小鼠在ATM和（II）CBX1条件敲除小鼠的情况下（I）CBX1 +/-小鼠的影响。这些研究将定义HP1B调节细胞IR反应和肿瘤发生的机制。我们的假设 - 非启用修饰因子HP1B调节染色质结构，并通过与DDR成分的相互作用，导致过度发生 - 是一种新颖的思想，需要深度研究。对正常组织和肿瘤组织染色质结构之间生化差异的机械基础的进一步理解将有助于开发新的策略，以改变IR反应并改善临床放射治疗。