Minority Predoctoral Fellowship Program

少数族裔博士前奖学金计划

• 批准号: 7112438
• 负责人: BRENDA SALERNO
• 金额: $ 4.35万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7112438
• 关键词: DNA repair; cellulose; enzyme activity; genetic regulation; inositol phosphates; ion exchange chromatography; mass spectrometry; molecular biology information system; molecular cloning; molecular genetics; phosphoester ligase; predoctoral investigator; protein protein interaction; protein purification; protein sequence; protein structure function; recombinant proteins; serine threonine protein kinase; surface plasmon resonance

DESCRIPTION (provided by applicant): Non-Homologous End-Joining (NHEJ) is a DNA double strand break repair pathway essential in maintaining the integrity of the genome. NHEJ requires the DNA Ligase IV/XRCC4 complex and the DNA-dependent Protein Kinase (DNA-PK), which is formed by DNA-PKcs, and the heterodimer Ku70/80. Inositol hexakisphosphate was shown to stimulate NHEJ in vitro, however, in vitro studies also suggest that NHEJ can be further stimulated by one or more additional unidentified factors. This research project will investigate the participation of an additional stimulatory factor in mammalian NHEJ. First, the unidentified stimulatory activity will be isolated by fractionation of human whole cell Extracts. Second, candidate stimulatory factors will be identified using protein sequencing and database searches, and participation of each candidate stimulatory factor will be confirmed utilizing the in vitro end-joining assay. Finally, the mechanism of NHEJ stimulation by the discovered factor will be explored. Results from this research will help understand the mechanism of NHEJ in vivo. This will facilitate the development of therapies for people with mutated NHEJ genes, and also anticancer treatments that can target NHEJ factors to sensitize cells before radiotherapy.

描述（由申请人提供）：非同源最终连接（NHEJ）是DNA双链断裂修复途径，对于维持基因组的完整性必不可少。 NHEJ需要由DNA-PKCS形成的DNA连接酶IV/XRCC4复合物和DNA依赖性蛋白激酶（DNA-PK）和Heterodimer KU70/80。肌醇六磷酸磷酸盐被证明可以在体外刺激NHEJ，但是，体外研究还表明，一个或多个其他未鉴定的因素可以进一步刺激NHEJ。该研究项目将调查附加刺激因子在哺乳动物NHEJ中的参与。首先，未鉴定的刺激活性将通过人类全细胞提取物的分馏来分离。其次，将使用蛋白质测序和数据库搜索来鉴定候选刺激因素，并使用体外最终结合测定法确认每个候选刺激因子的参与。最后，将探讨通过发现因子的NHEJ刺激机制。这项研究的结果将有助于了解体内NHEJ的机制。这将促进为NHEJ基因突变的人的疗法开发，以及可以针对NHEJ因素以在放射治疗之前敏化细胞的抗癌治疗方法。