NuMA function in nonmalignant and malignant breast cells

NuMA 在非恶性和恶性乳腺细胞中的功能

基本信息

批准号：
8514778
负责人：
SOPHIE A. LELIEVRE
金额：
$ 2.91万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-09-26 至 2013-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8514778
关键词：
Actins Acute Promyelocytic Leukemia Affect Affinity Chromatography Amino Acid Sequence Antibodies Architecture Area Behavior Binding Biochemistry Blocking Antibodies Breast Cancer Control Cell Cycle Regulation Cell Nucleus Cell Proliferation Cells Cellular Structures Characteristics Chimeric Proteins Chromatin Chromatin Remodeling Factor Chromatin Structure Chromatography Co-Immunoprecipitations Complex Data Development Disease Distal Environment Epithelial Epithelial Cells Equilibrium Euchromatin Event Extracellular Matrix GAS41 gene Gene Expression Gene Expression Regulation Genes Genetic Transcription Goals Heterochromatin Histones ISWI Indium Interphase Link Location Malignant - descriptor Malignant Neoplasms Mammary gland Mediating Medical Mitotic Spindle Apparatus Mitotic spindle Modeling Molecular Multiprotein Complexes Mutate Non-Malignant Nuclear Nuclear Proteins Peptides Phenotype Positioning Attribute Premalignant Proteins Regulation Relative (related person)Research Research Personnel Role Shapes Signal Pathway Structural Protein System Testing Therapeutic Therapeutic Intervention Time Tretinoin Work base calponin cancer cell chromatin remodeling genetic regulatory protein high risk histone modification innovation leukemia malignant breast neoplasm member neutrophil novel prevent promoter tumor tumor progression

项目摘要

DESCRIPTION (provided by applicant): Alterations in chromatin structure and gene expression are a hallmark of cancer. Proteins involved in chromatin remodeling complexes (CRCs) directly affect chromatin structure by modifying DMA and histones, and CRCs targeting is altered in cancer. Structural proteins like NuMA, considered as organizers of nuclear architecture, also interact with chromatin, and their distribution is altered in cancer. However, the role of NuMA in the control of gene expression is unknown. NuMA co-isolates with components of ATP-dependent CRCs. Altering NuMA using an antibody against its C-terminus (CT) or by expressing the distal portion of its CT modifies chromatin structure and the phenotype of breast epithelial cells. CT-truncated NuMA and mutated NuMA are implicated in leukemia and breast cancer development, respectively. We propose to investigate the mechanisms by which NuMA controls chromatin structure and gene expression in breast epithelial cells. The hypothesis is that NuMA participates in the control of gene transcription by interacting with components of CRCs, in order to target these complexes to specific genes. Three aims are proposed: (1) To identify the specific CRCs in which NuMA is involved by assessing the interaction of NuMA with components of SNF2h and BAF types of ATP-dependent CRCs by affinity chromatography and co- immunoprecipitation, and assessing the effect of inhibiting NuMA expression on the assembly and function of CRCs; (2) To analyze the primary sequence of NuMA with regards to its involvement in chromatin regulation by comparing the effects of interrupting the function of HPC2-like, GAS41-binding, and CH-binding regions of NuMA on chromatin organization, CRC function, and mammary epithelial differentiation; (3) To define the role of NuMA in gene expression control by position effect, by interfering with NuMA function and evaluating the expression status (on or off) of genes that control cell proliferation in relation to their localization and the presence of NuMA-containing CRCs at their promoter. Significance: A current scientific and medical challenge is to unravel the mechanisms that underlie changes in the machinery or regulation of gene expression during cancer development. Understanding these mechanisms will help develop strategies to prevent cancer progression and control tumor behavior. Thus, it is time to emphasize the role of structural nuclear proteins in gene transcription in order to get a complete picture of gene expression control.

描述（由申请人提供）：染色质结构和基因表达的改变是癌症的标志。参与染色质重塑复合物（CRC）的蛋白质通过修饰DMA和组蛋白直接影响染色质结构，并且靶向CRCS的靶向改变了癌症。像Numa这样的结构蛋白，被认为是核结构的组织者，也与染色质相互作用，并且它们的分布在癌症中发生了改变。但是，NUMA在基因表达的控制中的作用尚不清楚。 NUMA与依赖ATP的CRC的组成部分共叠。使用对其C末端（CT）的抗体改变NUMA，或通过表达其CT的远端部分修饰染色质结构和乳腺上皮细胞的表型。 CT截断的NUMA和突变的NUMA分别与白血病和乳腺癌发育有关。我们建议研究NUMA控制乳腺上皮细胞中染色质结构和基因表达的机制。假设是，Numa通过与CRC的成分相互作用，以将这些复合物靶向特定基因来控制基因转录。提出了三个目的：（1）通过评估NUMA与SNF2H和BAF类型的ATP依赖性CRC的相互作用来确定NUMA参与的特定CRC，并通过亲和力色谱和共同沉淀，并评估抑制抑制抑制效果。 CRC的组装和功能上的NUMA表达；（2）通过比较中断类似于HPC2的功能的NUMA的主要序列与其参与染色质调节的序列有关乳腺上皮分化；（3）通过干扰NUMA功能并评估控制细胞增殖与其定位和含Numa的存在的基因的表达状态（ON或OFF）来定义NUMA在基因表达控制中的作用，并评估基因的表达状态（ON或OFF） CRC的发起人。意义：当前的科学和医学挑战是揭示癌症发育过程中基因表达的机制或调节的机制。了解这些机制将有助于制定防止癌症进展并控制肿瘤行为的策略。因此，现在是时候强调结构性核蛋白在基因转录中的作用，以便获得基因表达控制的完整图片。