Dissecting BRCA1-PALB2 Activity in DNA Repair and Development

剖析 BRCA1-PALB2 在 DNA 修复和发育中的活性

基本信息

批准号：
10664883
负责人：
Neil Johnson
金额：
$ 38.66万
依托单位：
RESEARCH INST OF FOX CHASE CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-05 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10664883
关键词：
Alleles Amino Acids BRCA1 Mutation BRCA1 Protein BRCA1 gene BRCA2 Mutation BRCA2 gene Binding Biochemical Biological Assay Breast Cells Cessation of life Characteristics Chromatin Coiled-Coil Domain Complex DNA DNA Damage DNA Double Strand Break DNA Repair DNA Repair Pathway Defect Development Disease Embryo Embryonic Development Experimental Genetics Fanconi&apos s Anemia Filament Genetic Epistasis Genomic Instability Goals Health Heterodimerization Heterozygote Human Cell Line Immunoprecipitation Kinetics Knock-out Laboratories Link Maintenance Malignant Neoplasms Malignant neoplasm of ovary Mediating Molecular Mus Mutant Strains Mice Mutation Nature PALB2 gene Patients Peptides Phenotype Proteins Role Site Structure System Tertiary Protein Structure United States Work developmental disease experimental study genome integrity homologous recombination insight malignant breast neoplasm mouse genetics mouse model mutant mouse model mutation carrier neonatal mice novel postnatal development prevent protein complex recruit ubiquitin ligase ubiquitin-protein ligase

项目摘要

PROJECT SUMMARY The BRCA1 and PALB2 proteins directly heterodimerize through their respective coiled-coil (CC) domains, facilitating the formation of a larger BRCA1-PALB2-BRCA2-RAD51 complex that is required for RAD51 filament formation. Currently, little is known about the structural basis of the BRCA1-PALB2 interaction, including the residue alignment and orientation of physically interacting -helices. Elucidating CC molecular interactions is critical for understanding how mutations found in patients disrupt peptide interactions and promote disease. Protein CC domains are capable of mediating interactions with several CC domain containing partners. However, BRCA1 and PALB2 form the only known interaction that is mediated by their respective CC domains. Whether BRCA1 and PALB2 CC domains exclusively interact with one another, or if there are additional CC interactions and functions is unknown. Our laboratory has developed novel BRCA1 and PALB2 CC domain mutant mouse models to investigate the significance of this interaction in DNA repair and organismal health. We have also purified CC peptides so that biochemical assays can be performed assessing the effects of mutations on complex interactions and activity. The only known function of BRCA1 and PALB2 CC domains is to interact with one another, thus, BRCA1CC and PALB2CC homozygous mice might be expected to have identical phenotypes. However, while BRCA1CC mice are born at sub-Mendelian ratios and neo-natal mice demonstrate a range of developmental defects, PALB2CC homozygosity resulted in early embryonic lethality. Because BRCA1CC and PALB2CC mice have distinct phenotypes, we hypothesize that CC domains facilitate protein interactions beyond the BRCA1-PALB2 heterodimer that promote DNA repair and embryonic development. We will address the following Specific Aims: 1) Determine biochemical CC interactions and activity; 2) Uncover DNA repair and developmental defects in CC mutant mice; and 3) Elucidate mechanisms of BRCA1-PALB2 complex recruitment to DNA breaks. Collectively, the proposed experiments will yield new insight into the mechanism by which the BRCA1-PALB2 complex protects from genome instability.

项目摘要 BRCA1和PALB2蛋白通过其各自的盘绕螺旋（CC）域直接异二聚二聚体，促进Rad51细丝所需的较大BRCA1-PALB2-BRCA2-RAD51复合物的形成形成。当前，关于BRCA1-PALB2相互作用的结构基础知之甚少，包括物理相互作用的残留对齐和方向。阐明CC分子相互作用是对于了解患者中发现的突变如何破坏肽相互作用并促进疾病至关重要。蛋白质CC结构域能够介导与含有伴侣的多个CC结构域的相互作用。然而， BRCA1和PALB2构成了由其各自的CC域介导的唯一已知相互作用。无论 BRCA1和PALB2 CC域专门相互作用，或者是否还有其他CC相互作用功能是未知的。我们的实验室开发了新颖的BRCA1和PALB2 CC域突变小鼠研究这种相互作用在DNA修复和有机健康中的重要性的模型。我们也有纯化的CC Pepperides，以便可以评估突变对复合物的影响的生化测定相互作用和活动。 BRCA1和PALB2 CC域的唯一已知功能是与一个因此，另一种可能会有望具有相同的表型。但是，尽管BRCA1CC小鼠以亚孟德尔的比率出生，而新生儿小鼠则表现出一系列发育缺陷，PALB2CC纯合性导致早期胚胎致死性。因为brca1cc和 PALB2CC小鼠具有不同的表型，我们假设CC域喜欢的蛋白质相互作用以外促进DNA修复和胚胎发育的BRCA1-PALB2异二聚体。我们将解决以下特定目的：1）确定生化CC相互作用和活性； 2）发现DNA修复和 CC突变小鼠的发育缺陷； 3）阐明BRCA1-PALB2复合募集机制到DNA断裂。总的来说，拟议的实验将对该机制产生新的见解 BRCA1-PALB2复合物可保护基因组不稳定性。

项目成果

期刊论文数量（8）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Replication gaps are a key determinant of PARP inhibitor synthetic lethality with BRCA deficiency.

DOI：
10.1016/j.molcel.2021.06.011
发表时间：
2021-08-05
期刊：
MOLECULAR CELL
影响因子：
16
作者：
Cong, Ke;Peng, Min;Kousholt, Arne Nedergaard;Lee, Wei Ting C.;Lee, Silviana;Nayak, Sumeet;Krais, John;VanderVere-Carozza, Pamela S.;Pawelczak, Katherine S.;Calvo, Jennifer;Panzarino, Nicholas J.;Turchi, John J.;Johnson, Neil;Jonkers, Jos;Rothenberg, Eli;Cantor, Sharon B.
通讯作者：
Cantor, Sharon B.

BRCA1 Mutations in Cancer: Coordinating Deficiencies in Homologous Recombination with Tumorigenesis.

DOI：
10.1158/0008-5472.can-20-1830
发表时间：
2020-11-01
期刊：
Cancer research
影响因子：
11.2
作者：
Krais JJ;Johnson N
通讯作者：
Johnson N

Brca1 mutations in the coiled-coil domain impede Rad51 loading on DNA and mouse development.

卷曲螺旋结构域中的 Brca1 突变阻碍了 Rad51 在 DNA 上的加载和小鼠发育。

DOI：
10.1080/23723556.2020.1786345
发表时间：
2020
期刊：
Molecular & cellular oncology
影响因子：
2.1
作者：
Krais,JJ;Johnson,N
通讯作者：
Johnson,N

BRCA1 secondary splice-site mutations drive exon-skipping and PARP inhibitor resistance.

BRCA1 二次剪接位点突变导致外显子跳跃和 PARP 抑制剂耐药。

DOI：
10.1101/2023.03.20.23287465
发表时间：
2023
期刊：
medRxiv : the preprint server for health sciences
影响因子：
0
作者：
Nesic,Ksenija;Krais,JohnJ;Vandenberg,CassandraJ;Wang,Yifan;Patel,Pooja;Cai,KathyQ;Kwan,Tanya;Lieschke,Elizabeth;Ho,Gwo-Yaw;Barker,HollyE;Bedo,Justin;Casadei,Silvia;Farrell,Andrew;Radke,Marc;Shield-Artin,Kristy;Penington,
通讯作者：
Penington,

BRCA1 Haploinsufficiency Is Masked by RNF168-Mediated Chromatin Ubiquitylation.

BRCA1 单倍体不足被 RNF168 介导的染色质泛素化掩盖。

DOI：
10.1016/j.molcel.2018.12.010
发表时间：
2019
期刊：
Molecular cell
影响因子：
16
作者：
Zong,Dali;Adam,Salomé;Wang,Yifan;Sasanuma,Hiroyuki;Callén,Elsa;Murga,Matilde;Day,Amanda;Kruhlak,MichaelJ;Wong,Nancy;Munro,Meagan;RayChaudhuri,Arnab;Karim,Baktiar;Xia,Bing;Takeda,Shunichi;Johnson,Neil;Durocher,Daniel;Nusse
通讯作者：
Nusse