Structural and Functional Characterization of BRCA1/BARD1

BRCA1/BARD1 的结构和功能表征

基本信息

批准号：
8309249
负责人：
Rachel E Klevit
金额：
$ 50.97万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
1998
资助国家：
美国
起止时间：
1998-12-21 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8309249
关键词：
Affinity Ankyrin Repeat BARD1 gene BRCA1 gene Binding Binding Proteins Biochemical Biological Biological Assay C-terminal Cell physiology Cells Chromosome abnormality Complex Crystallography DNA Damage Enzymes Estrogen Receptors Event Functional disorder Future Genes Genome Stability Goals Grant Health Human Human Ubiquitin In Vitro Inherited Investigation Knock-in Mouse Knock-out Lead Learning Ligase Link Maintenance Malignant Neoplasms Malignant neoplasm of ovary Mass Spectrum Analysis Methods Molecular Mono-S Multienzyme Complexes Mus Mutation N-terminal Nature Outcome Play Polyubiquitin Progesterone Receptors Protein Binding Proteins Regulation Resolution Role Substrate Interaction System Techniques Testing Tumor Suppressor Proteins Ubiquitin Ubiquitin-Conjugating Enzymes Ubiquitination Woman Work Yeasts in vivo insight lifetime risk malignant breast neoplasm mutant protein complex research study response three dimensional structure tool ubiquitin ligase ubiquitin-protein ligase yeast two hybrid system

项目摘要

The breast and ovarian cancer tumor suppressor protein, BRCA1, and its obligate protein partner, BARD1 are critical to proper functioning of fundamental cellular processes that support genomic stability. To date, a single biochemical activity has been identified for BRCA1/BARD1, namely, they function together as a ubiquitin E3 ligase. Although it is believed that the ability of BRCA1/BARD1 to modify specific cellular proteins with ubiquitin is fundamental to its role as a tumor suppressor, the details and ramifications of this relationship remain to be elucidated. During the past grant period, we discovered that BRCA1/BARD1 can function with ten human ubiquitin-conjugating enzymes (E2s) and that the ultimate product generated by BRCA1/BARD1 on substrates depends on the E2 present. We also identified several new substrates, including the estrogen receptor. In the next grant period we will expand our focus to include the essential subunit, BARD1, more explicitly and will participate in a collaborative effort to develop new molecular insights and tools for another RING E3 ligase that is critical in cancers, Mdm2/MdmX. The overall goals for the next grant period are 1) elucidate structural and functional determinants of mono-ubiquitin transfer and poly-ubiquitin chain formation by BRCA1/BARD1 and its interacting E2s, 2) investigate the molecular and structural determinants of BRCA1/BARD1-substrate interactions, 3) characterize BARD1 and its interactions, and 4) identify the human E2s that interact with Mdm2/MdmX, the ligase responsible for p53 regulation in vivo. A broad experimental approach will be undertaken, including biochemical, structural, molecular biological, and cellular techniques. Results from these studies will provide new insights into BRCA1/BARD1 function and will contribute to the general understanding of protein ubiquitination.

乳腺癌和卵巢癌肿瘤抑制蛋白 BRCA1 及其专性蛋白伴侣， BARD1 对于支持基因组的基本细胞过程的正常运行至关重要稳定。迄今为止，已鉴定出 BRCA1/BARD1 的单一生化活性，即它们作为泛素 E3 连接酶一起发挥作用。尽管人们相信 BRCA1/BARD1 的能力用泛素修饰特定的细胞蛋白是其作为肿瘤抑制因子的基础这种关系的细节和后果仍有待阐明。在过去的资助期内，我们发现 BRCA1/BARD1 可以与十种人类泛素结合酶 (E2) 一起发挥作用，并且 BRCA1/BARD1 在底物上产生的最终产物取决于 E2 的存在。我们还发现了几种新的底物，包括雌激素受体。在下一个资助期内，我们将将我们的重点扩大到更明确地包括重要的子单元 BARD1，并将参与共同努力为另一种 RING E3 连接酶开发新的分子见解和工具 Mdm2/MdmX 对癌症至关重要。下一个资助期的总体目标是 1) 阐明结构单泛素转移和多泛素链形成的功能决定因素 BRCA1/BARD1 及其相互作用的 E2，2) 研究 BRCA1/BARD1 的分子和结构决定因素 BRCA1/BARD1-底物相互作用，3) 表征 BARD1 及其相互作用，4) 识别与 Mdm2/MdmX 相互作用的人类 E2，Mdm2/MdmX 是负责体内 p53 调节的连接酶。广泛的将采用实验方法，包括生物化学、结构、分子生物学和细胞技术。这些研究的结果将为 BRCA1/BARD1 功能提供新的见解并将有助于对蛋白质泛素化的一般理解。