The Role of 53BP1 Interacting Proteins in Regulation of DNA Repair Choice
53BP1 相互作用蛋白在 DNA 修复选择调节中的作用
基本信息
- 批准号:8916643
- 负责人:
- 金额:$ 3.32万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2013
- 资助国家:美国
- 起止时间:2013-09-16 至 2016-09-15
- 项目状态:已结题
- 来源:
- 关键词:Animal ModelBRCA1 geneBiological AssayBreastCandidate Disease GeneCell CycleCell Cycle CheckpointCell LineCellsCisplatinComplexDNADNA Double Strand BreakDNA RepairDNA Repair PathwayDNA repair proteinDataDefectExcisionFunctional disorderGenomic InstabilityGenomicsHealthHumanImmunohistochemistryInheritedLeadMalignant NeoplasmsMalignant neoplasm of ovaryMeasuresMediatingModalityModelingMusMutationNonhomologous DNA End JoiningNuclear Matrix-Associated ProteinsOvarianPathway interactionsPhasePhenotypePlayProteinsProteomicsRecruitment ActivityRegulationResistanceReverse Transcriptase Polymerase Chain ReactionRoleS PhaseScaffolding ProteinSet proteinSiteSmall Interfering RNATumor Suppressor ProteinsWomanbasecancer cellcrosslinkdesignfunctional losshigh riskhomologous recombinationhuman H2AX proteininhibitor/antagonistinnovationinsightmalignant breast neoplasmmutantnovelp53-binding protein 1protein protein interactionrepairedrestoration
项目摘要
DESCRIPTION (provided by applicant): 53BP1 is a nuclear scaffolding protein and is involved in DNA repair and cell cycle checkpoint control. Upon induction of DNA double strand breaks, 53BP1 is rapidly phosphorylated by ATM/ATR, and recruited to sites of DNA breaks where it co-localizes with phosphorylated histone H2AX and other DNA repair proteins such as BRCA1. Women with functional loss of BRCA1 are at significantly higher risk for developing breast or ovarian malignancies. Cancer cells that are deficient in BRCA1 have significant homologous recombination mediated DNA repair (HDR) defect, genomic instability, and sensitivity to PARP inhibitors (PARPi) and cisplatin. The current model is that cells with BRCA1 deficiency have unconstrained 53BP1 activity. 53BP1, by yet uncharacterized mechanism, inhibits end resection at DNA double strand breaks and thus, promoting non- homologous end joining (NHEJ) and suppressing HDR. Interestingly, cells with both BRCA1 and 53BP1 deficiencies have partial restoration of HDR and resistance to PARPi and cisplatin. These observations have given rise to notion that 53BP1 and BRCA1 are critical regulators of DNA repair choice. The mechanism, however, by which 53BP1 executes its function in inhibiting end-resection and HDR, and promoting NHEJ is completely unknown. 53BP1 has no known enzymatic function, and is thought to function as a scaffold protein that acts as a platform for protein-protein interactions.
We propose that 53BP1 interacts with a set of proteins that are necessary for its role in inhibiting end resection. These interactions are critical for the suppression of HDR seen in BRCA1-/- cells. Loss of 53BP1 interacting proteins will restore HDR in BRCA1 -/- cells and lead to chemoresistance. Recent studies have identified RAD50, a component of MRN complex, and TopBP1, a regulator of CtIP, as 53BP1 interacting proteins. As the MRN complex and CtIP plays critical role in end resection and HDR, it is plausible that 53BP1 mediates the DNA repair phenotype in Brca1 - /- cells via RAD50 and TopBP1. siRNA knockdown will be performed on 53BP1 interacting proteins in Brca1 -/- cell line to measure the restoration of HDR and resistance to PARPi and cisplatin. Furthermore, we will use unbiased high-throughput proteomics approach to identify novel 53BP1-interacting proteins in both Brca1 +/+ and Brca1-/- cell lines. Role of these candidate genes will be analyzed in DNA repair using specific GFP-based assays for HR and NHEJ. Expression of known and novel 53BP1 interacting proteins in human and murine BRAC1-related cancers will be determined using immunohistochemistry, RT-PCR, and genomic sequencing. The ultimate aim of this proposal is to gain new insight into how 53BP1 functions to regulate DNA repair choice in both BRCA1-mutant and BRCA1-wt cells. This will lead to new understanding of how DNA repair pathways are regulated as well as innovative approaches to target chemo resistance in human cancers.
描述(由申请人提供):53BP1是一种核支架蛋白,参与DNA修复和细胞周期检查点控制。在DNA双链断裂诱导下,53BP1被ATM/ATR迅速磷酸化,并招募到DNA断裂位点,与磷酸化组蛋白H2AX和其他DNA修复蛋白(如BRCA1)共定位。BRCA1功能缺失的女性患乳腺或卵巢恶性肿瘤的风险明显更高。缺乏BRCA1的癌细胞具有显著的同源重组介导的DNA修复(HDR)缺陷、基因组不稳定性以及对PARP抑制剂(PARPi)和顺铂的敏感性。目前的模型是具有BRCA1缺陷的细胞具有不受约束的53BP1活性。53BP1抑制DNA双链断裂末端切除,从而促进非同源末端连接(non- homologous end joining, NHEJ),抑制HDR,机制尚不明确。有趣的是,BRCA1和53BP1缺陷的细胞都能部分恢复HDR,并对PARPi和顺铂产生耐药性。这些观察结果表明,53BP1和BRCA1是DNA修复选择的关键调节因子。然而,53BP1在抑制末端切除和HDR以及促进NHEJ中的作用机制是完全未知的。53BP1没有已知的酶功能,被认为是一种支架蛋白,作为蛋白质相互作用的平台。
项目成果
期刊论文数量(0)
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Jinesh Shailesh Gheeya其他文献
Jinesh Shailesh Gheeya的其他文献
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{{ truncateString('Jinesh Shailesh Gheeya', 18)}}的其他基金
The Role of 53BP1 Interacting Proteins in Regulation of DNA Repair Choice
53BP1 相互作用蛋白在 DNA 修复选择调节中的作用
- 批准号:
8786396 - 财政年份:2013
- 资助金额:
$ 3.32万 - 项目类别:
The Role of 53BP1 Interacting Proteins in Regulation of DNA Repair Choice
53BP1 相互作用蛋白在 DNA 修复选择调节中的作用
- 批准号:
8526829 - 财政年份:2013
- 资助金额:
$ 3.32万 - 项目类别:
The Role of 53BP1 Interacting Proteins in Regulation of DNA Repair Choice
53BP1 相互作用蛋白在 DNA 修复选择调节中的作用
- 批准号:
9119797 - 财政年份:2013
- 资助金额:
$ 3.32万 - 项目类别:
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