Ubiquitin-mediated regulation of the RAD51D homologous recombination protein

泛素介导的 RAD51D 同源重组蛋白调节

基本信息

批准号：
8689271
负责人：
DOUGLAS Lee PITTMAN
金额：
$ 32.53万
依托单位：
UNIVERSITY OF SOUTH CAROLINA AT COLUMBIA
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2018-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8689271
关键词：
Alternative Splicing Amino Acids Antineoplastic Agents BRCA1 gene BRCA2 gene Biology Biomedical Research Cancer-Predisposing Gene Carcinogenesis Mechanism Cellular biology Chromosomal Stability Clinic Co-Immunoprecipitations Complex Confocal Microscopy Congenital Abnormality DNA DNA Damage DNA Interstrand Crosslinking Data Defect Development Doctor of Pharmacy Double Strand Break Repair Drug Targeting Enzymes Exposure to Family member Fellowship Genes Genetic Genome Genomic Instability Human Genome Individual Infertility Institution Laboratories Lysine Malignant Neoplasms Malignant neoplasm of ovary Mass Spectrum Analysis Mediating Minority Mutation New York Pathway interactions Pharmacologic Substance Pharmacy Students Phenotype Post-Translational Protein Processing Premature aging syndrome Process Program Reviews Protein Isoforms Proteins Rad51 recombinase Recruitment Activity Regulation Research Research Training Science Scientist Site South Carolina Techniques Testing Texas Training Ubiquitin Ubiquitination Underrepresented Minority Universities Yeasts cancer cell chemotherapy college demographics environmental agent experience graduate student high school homologous recombination member multicatalytic endopeptidase complex mutant novel programs protein function public health relevance repaired research study response teacher tool ubiquitin-protein ligase undergraduate student yeast two hybrid system

项目摘要

PROJECT SUMMARY Mutations in RAD51 genes confer genome instability phenotypes associated with infertility, birth defects, and cancer. Each of the RAD51 family members are central components of the error-free double strand break (DSB) repair pathway known as homologous recombination (HR), and RAD51D was recently identified as an ovarian cancer susceptibility gene. Even though it is known that HR defects predispose individuals to malignancy and confers cancer cell sensitivity to DNA damaging chemotherapy agents, single-protein analyses have revealed little functional information. Therefore, we performed RAD51D protein interaction screens and identified RNF138, a novel RING domain E3 ubiquitin ligase enzyme. Ubiquitin is a 76-amino acid protein that modifies protein functions or marks them for proteasome degradation, leading to the central hypothesis of this application: RAD51D forms adaptive complexes that are tightly regulated by RNF138 to process DNA breaks and maintain genome integrity. To test this hypothesis, the first aim will determine how RNF138 and RAD51D interaction regulates the HR DNA damage response pathway. The second aim will identify the ubiquitin linkage arrangements and sites along the RAD51D protein and their association with DNA damage response. Over the course of this project, undergraduate, graduate, and Doctor of Pharmacy students will receive outstanding biomedical research training. Undergraduate students are recruited from the highly selective University of South Carolina Honors College, which is ranked first by the Public University Honors Program "Review of Fifty Public University Honors Programs." Therefore, investigating RAD51D post-translational modifications will uncover mechanisms of carcinogenesis pathways and identify potential cancer drug targets as well as provide training in state-of-the-art biomedical science techniques.

项目摘要 RAD51基因的突变赋予基因组不稳定性表型与不育症，先天缺陷和癌症。每个RAD51家庭成员都是无错误的双链断裂的中心组成部分（DSB）修复途径称为同源重组（HR），而RAD51D最近被确定为卵巢癌敏感性基因。即使知道人力资源缺陷使人倾向于恶性肿瘤并赋予癌细胞对DNA损害化疗剂的敏感性，单蛋白分析揭示了几乎没有功能信息。因此，我们进行了RAD51D蛋白相互作用筛选和鉴定出RNF138，这是一种新型的环域E3泛素连接酶。泛素是一种76-氨基酸蛋白修饰蛋白质功能或标记它们以降解蛋白酶体，从而导致中心假设应用：RAD51D形成由RNF138严格调节以处理DNA断裂的自适应复合物并保持基因组完整性。为了检验这一假设，第一个目标将决定RNF138和RAD51D如何相互作用调节HR DNA损伤响应途径。第二个目标将确定泛素连接沿RAD51D蛋白的排列和位点及其与DNA损伤反应的关联。在这个项目的课程，本科，研究生和药房的学生将获得出色的生物医学研究培训。本科生是从高度选择性大学招募的南卡罗来纳州荣誉学院，该学院由公立大学荣誉计划排名第一。公立大学荣誉计划。”因此，调查RAD51D翻译后修改将发现癌变途径的机制，并确定潜在的癌症药物靶标，并提供在最先进的生物医学科学技术中培训。