Minority Predoctoral Fellowship Program

少数族裔博士前奖学金计划

• 批准号: 7119950
• 负责人: Roberto A Neisa
• 金额: $ 3.63万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-27 至 2007-07-26
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7119950
• 关键词: SDS polyacrylamide gel electrophoresis; acute myelogenous leukemia; carcinogenesis; cell differentiation; cell growth regulation; cell line; cyclin dependent kinase; cyclins; density gradient ultracentrifugation; enzyme activity; flow cytometry; immunoprecipitation; neoplastic cell; phosphorylation; predoctoral investigator; protein binding; protein localization; protein protein interaction; western blottings

DESCRIPTION (provided by applicant): Human cyclin A1 is expressed at its highest levels in spermatocytes but also in leukemic cell lines and leukemic cells obtained from patients with acute myeloid leukemia (AML), particularly acute promyelocytic leukemia (APL). Interestingly, in mouse leukemia models and in preliminary investigations on the human APL cell line NB4, the localization of cyclin A1 is predominantly cytoplasmic, unlike its nuclear localization in germ cells. The hypothesis that will be tested is that the cytoplasmic accumulation of cyclin A1 contributes to its transforming potential. NB4 cells will be examined for cell cycle-specific cyclin A1 subcellular localization patterns. The cytoplasmic distribution of cyclin A1 will be determined by sub-cellular fractionation followed by immunoblotting. Coimmunoprecipitation assays will be used to determine the binding partners of cyclin A1, including its partner kinase(s) and substrates. Kinase assays will be performed to evaluate the activity of cyclin A1-CDK complexes. Finally, the contribution of cytoplasmic cyclin A1 localization to the transforming phenotype will be examined by fusing cytoplasmic organelle-specific targeting sequences to cyclin A1 constructs.

描述（由申请人提供）：人细胞周期蛋白A1在精子细胞中以最高水平表达，以及从急性髓样白血病（AML）患者获得的白血病细胞系和白血病细胞中，尤其是急性临床前临床前细胞性白血病（APL）。有趣的是，在小鼠白血病模型和对人APL细胞系NB4的初步研究中，细胞周期蛋白A1的定位主要是细胞质的，这与其在生殖细胞中的核定位不同。将要检验的假设是细胞周期蛋白A1的细胞质积累有助于其转化潜力。 NB4细胞将检查细胞周期特异性细胞周期A1亚细胞定位模式。细胞周期蛋白A1的细胞质分布将通过亚细胞分级，然后通过免疫印迹来确定。共免疫沉淀测定将用于确定细胞周期蛋白A1的结合伙伴，包括其伴侣激酶和底物。激酶测定将进行评估细胞周期蛋白A1-CDK复合物的活性。最后，通过将细胞质细胞器特异性靶向序列融合到Cyclin A1构建体的细胞质环蛋白A1定位对转化表型的贡献。