Ubiquitin-mediated regulation of the RAD51D homologous recombination protein

项目总结 RAD51基因突变使基因组不稳定表型与不孕症、出生缺陷和 癌症。RAD51家族的每个成员都是无错误双链断裂的中心组件 (DSB)修复途径称为同源重组(HR)，RAD51D最近被发现是一种 卵巢癌易感基因。即使我们知道人力资源缺陷会使个人倾向于 单蛋白分析显示，肿瘤细胞对破坏DNA的化疗药物具有敏感性 几乎没有透露什么功能信息。因此，我们进行了RAD51D蛋白质相互作用筛选和 鉴定了一种新的环区E3泛素连接酶RNF138。泛素是一种由76个氨基酸组成的蛋白质 修饰蛋白质功能或将其标记为蛋白酶体降解，导致了这一中心假说 应用：RAD51D形成受RNF138严格调控的适应性复合体，处理DNA断裂 并保持基因组的完整性。为了检验这一假设，第一个目标将确定RNF138和RAD51D如何 相互作用调节HR DNA损伤反应通路。第二个目标是确定泛素的连接 RAD51D蛋白的排列和位置及其与DNA损伤反应的关系。超过了 本项目课程，本科生、研究生和药学博士研究生将获得优异成绩 生物医学研究培训。本科生从高度挑剔的伊利诺伊大学招收 南卡罗来纳州荣誉学院，在公立大学荣誉计划“五十年回顾”中排名第一 因此，调查RAD51D翻译后的修改将 揭示致癌途径的机制，识别潜在的抗癌药物靶点，并提供 接受最先进的生物医学科学技术培训。

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.