MOLECULAR CHARACTERIZATION OF MCLI PCNA INTERACTION

Fujise博士的长期目标是成为一名分子生物学家和心脏病专家，其主要重点是使用分子生物学方法预防和治疗心血管疾病。在Edward T.H. Yeh博士的指导下，Fujise博士开展了细胞凋亡调控领域的研究。在表征Bcl-2同源物MCL1（ML1髓样细胞白血病）的过程中，Fujise博士发现MCL1与PCNA（增殖细胞核抗原）之间存在一种新的相互作用。PCNA是一种29kDa的核蛋白，参与DNA修复和复制以及细胞增殖和分化。PCNA的作用与系统性红斑狼疮（SLE）、血管成形术后再狭窄等的发病机制有关。细胞凋亡及其缺失在癌症、艾滋病、心肌炎、动脉粥样硬化和心肌梗死等许多人类疾病中起着至关重要的作用。MCL1-PCNA相互作用可能是抗凋亡蛋白与细胞周期蛋白之间的重要分子联系。基于MCL1通过与PCNA的相互作用在DNA修复和复制以及细胞增殖和分化中发挥作用的假设，Fujise博士在此提出了MCL1-PCNA相互作用的特征。首先，Fujise博士将使用酵母双杂交系统进一步表征MCL1-PCNA相互作用的生化方面。其次，Fujise博士将研究MCL1-PCNA相互作用的功能意义，包括MCL1过表达对细胞增殖的影响，以及MCL1在PCNA存在下对DNA聚合酶delta、DNA甲基化酶和DNA修复酶的影响。最后，我们将评估细胞周期阶段、磷酸化和DNA损伤对MCL1- pcna相互作用和MCL1亚细胞定位的影响。在叶博士的指导下，Fujise博士将在人类疾病预防分子医学研究所的心血管疾病研究中心进行研究，将80%的专业时间用于研究。研究中心和研究所将为Fujise博士提供一个高度互动和富有成效的环境，以及他研究所需的所有设备。由包括导师在内的三名资深科学家组成的科学咨询委员会将指导藤濑博士的科学进步。完成本奖项后，Fujise博士将成为一名独立、高效的研究者，很有可能在细胞凋亡调控和心脏病预防和治疗领域做出重要贡献。

Dr. Fujise's long-term objectives is to serve as a molecular biologist and cardiologist, whose main focus is prevention and treatment of cardiovascular diseases using molecular biological approach. Under the guidance and direction of Dr. Edward T.H. Yeh, Dr. Fujise has performed the research in the field of apoptosis regulation. In the process of characterizing MCL1(ML1 myeloid cell leukemia), a Bcl-2 homologue, Dr. Fujise discovered a novel interaction between MCL1 and PCNA (Proliferating cell nuclear antigen). PCNA is a 29kDa nuclear protein involved in DNA repair and replication and cell proliferation and differentiation. The role of PCNA has been implicated in pathogenesis of SLE (Systemic lupus erythematosis), restenosis after angioplasty and others. Apoptosis as well as the lack of it play critical roles in many human diseases including cancer, AIDS, myocarditis, atherosclerosis and myocardial infarction. The MCL1-PCNA interaction may be an important molecular link between anti-apoptotic proteins and cell cycle proteins. Based on the hypothesis that MCL1 plays a role in DNA repair and replication and cell proliferation and differentiation through its interaction with PCNA, Dr. Fujise here proposes to characterize the MCL1-PCNA interaction over the five years of the Award. First, Dr. Fujise will further characterize biochemical aspects of MCL1-PCNA interaction, using yeast two hybrid system. Second, Dr. Fujise will investigate the functional significance of the MCL1-PCNA interaction, including the effect of MCL1 over- expression on cell proliferation and the effect of MCL1 on DNA polymerase delta, DNA methylase, and DNA repair enzymes in the presence of PCNA. Finally, the impact of cell cycle phases, phosphorylation and DNA damage on MCL1-PCNA interaction and MCL1's subcellular localization will be evaluated. With Dr. Yeh as his Mentor, Dr. Fujise will perform his research at Research Center for Cardiovascular Diseases within the Institute of Molecular Medicine for Prevention of Human Diseases, devoting 80 percent of his professional time to the research. The Research Center and Institute will provide Dr. Fujise with a highly interactive and productive environment as well as with all the equipment needed for his research. The Scientific Advisory Committee, consisting of three senior scientists including the mentor will guide Dr. Fujise's scientific progress. At the completion of this Award, Dr. Fujise will be an independent, highly productive investigator with a high likelihood of making critical contributions in the fieled of apoptosis regulations and in prevention and treatment of cardiac diseases.