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.

乳腺癌和卵巢癌抑制剂蛋白BRCA1及其强制性蛋白质伴侣， Bard1对于支持基因组的基本细胞过程的正常运作至关重要稳定。迄今作为泛素E3连接酶一起起作用。尽管相信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剂的作用至关重要的这种关系的细节和后果仍有待阐明。在过去的赠款期间，我们发现BRCA1/BARD1可以与十种人泛素结合酶（E2S）和 BRCA1/BARD1在底物上产生的最终产物取决于存在的E2。我们还确定了几种新的底物，包括雌激素受体。在下一个赠款期间，我们将将我们的重点扩大到更明确的基本亚基，Bard1，并将参与协作努力为另一个环RIGASE开发新的分子见解和工具癌症至关重要，MDM2/MDMX。下一个赠款期的总体目标是1）阐明结构以及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）研究的分子和结构决定因素 BRCA1/BARD1-SUBSTRATE相互作用，3）表征Bard1及其相互作用，4）确定与MDM2/MDMX相互作用的人E2，该连接酶负责体内p53调节。广泛将采用实验方法，包括生化，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。