ERAP140 - POSTTRANSLATIONAL MODIFICATION IDENTIFICATION

ERAP140 - 翻译后修饰鉴定

• 批准号: 6791941
• 负责人: TIMOTHY R GEISTLINGER
• 金额: $ 4.11万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-16 至 2007-04-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6791941
• 关键词: biochemistry; biological signal transduction; breast neoplasms; chromatin immunoprecipitation; drug resistance; estrogen receptors; gene expression; genetic regulation; mass spectrometry; microarray technology; neoplasm /cancer genetics; nuclear receptors; polymerase chain reaction; postdoctoral investigator; posttranslational modifications; protein protein interaction; protein structure function; proteomics; site directed mutagenesis; tamoxifen; transcription factor

DESCRIPTION (provided by applicant): Utilizing the latest techniques in genetics, biochemistry, and mass spectroscopy, I will focus on a recently discovered transcription factor, the estrogen receptor associated protein 140 (ERAP140) to determine its specific role in genomic regulation, identify and characterize posttranslational modifications (PTMs) of this 3rotein to evaluate their modulation of genomic regulation in healthy and diseased states. It is our hypothesis that the transcriptional activation potential of ERAP140 and its role in breast cancer, and tamoxifen resistant breast cancer, is likely modulated by, as yet, undetermined PTM states of ERAP140. I will focus on three 3reast tumor cell lines (MCF-7, T47D, and MDA231) with different pathologies expressing very high levels of ERAP140. Utilizing chromatin immunoprecipitation (CHIP), DNA promoter microarray analysis, isotopic labeling, and proteomics analysis of ERAP140 and its PTM state I will identify correlations with its altered pattern of promoter specific macromolecular transcription complex formation. Site-directed mutagenesis of the identified ERAP140 PTM sites will be evaluated in transcription translation assays and protein interaction assays for the mechanism of action behind these patterns.

描述（由申请人提供）：利用遗传学、生物化学和质谱方面的最新技术，我将重点关注最近发现的一种转录因子，即雌激素受体相关蛋白 140 (ERAP140)，以确定其在基因组调控中的具体作用，识别和表征这 3 个蛋白质的翻译后修饰 (PTM)，以评估其在健康和患病状态下对基因组调控的调节作用。我们的假设是，ERAP140 的转录激活潜力及其在乳腺癌和他莫昔芬耐药乳腺癌中的作用可能受到尚未确定的 ERAP140 PTM 状态的调节。我将重点关注三种 3reast 肿瘤细胞系（MCF-7、T47D 和 MDA231），它们具有不同的病理学，表达非常高水平的 ERAP140。利用染色质免疫沉淀 (CHIP)、DNA 启动子微阵列分析、同位素标记和 ERAP140 及其 PTM 状态 I 的蛋白质组学分析，将确定与其启动子特异性大分子转录复合物形成模式改变的相关性。已确定的 ERAP140 PTM 位点的定点诱变将在转录翻译测定和蛋白质相互作用测定中进行评估，以了解这些模式背后的作用机制。