MOLECULAR CHARACTERIZATION OF MCLI PCNA INTERACTION

MCLI PCNA 相互作用的分子表征

• 批准号: 6612747
• 负责人: Ken Fujise
• 金额: $ 12.47万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-07-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6612747
• 关键词: BCL2 gene /protein; DNA damage; DNA methylation; DNA repair; DNA replication; SDS polyacrylamide gel electrophoresis; apoptosis; binding sites; cell differentiation; cell growth regulation; myelogenous leukemia; phosphorylation; proliferating cell nuclear antigen; protein binding; protein localization; protein protein interaction; protein structure function; tissue /cell culture; transfection; yeast two hybrid system

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.

福吉斯博士的长期目标是作为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。 在爱德华·T.H.博士的指导和指导下Yeh，Fujise博士在凋亡调节领域进行了研究。 在表征MCL1（ML1髓样细胞白血病）的过程中，Bcl-2同源物，Fujise博士发现了MCL1和PCNA（增殖细胞核抗原）之间的新型相互作用。 PCNA是一种与DNA修复，复制以及细胞增殖和分化有关的29KDA核蛋白。 PCNA的作用与SLE的发病机理有关（全身性红斑狼疮），血管成形术后再狭窄等。 凋亡以及缺乏IT在许多人类疾病中起关键作用，包括癌症，艾滋病，心肌炎，动脉粥样硬化和心肌梗塞。 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。 基于以下假设：MCL1通过与PCNA的相互作用在DNA修复，复制和细胞增殖和分化中起作用，Fujise博士在此提议在奖项的五年内表征MCL1-PCNA相互作用。 首先，福吉斯博士将使用酵母两杂交系统进一步表征MCL1-PCNA相互作用的生化方面。其次，福吉斯博士将研究MCL1-PCNA相互作用的功能意义，包括MCL1过度表达对细胞增殖的影响以及MCL1对PCNA存在下DNA聚合酶三角洲，DNA甲基酶和DNA修复酶的影响。 最后，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。 富吉斯博士将以YEH为导师，将在预防人类疾病的分子医学研究所内的心血管疾病研究中心进行研究，将其专业时间的80％用于研究。 研究中心和研究所将为福吉斯博士提供高度互动和富有成效的环境以及他的研究所需的所有设备。 科学咨询委员会由包括导师在内的三名高级科学家组成，将指导福吉斯博士的科学进步。 该奖项结束后，福吉斯博士将是一位独立的高产研究者，很有可能在凋亡法规以及预防和治疗心脏疾病的情况下做出关键贡献。

项目成果

Embryonic lethality of fortilin-null mutant mice by BMP-pathway overactivation.

BMP 通路过度激活导致 fortilin 缺失突变小鼠的胚胎致死率。

• DOI: 10.1016/j.bbagen.2009.01.012
• 发表时间: 2009-05
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
• 影响因子: 3
• 作者: Koide, Yuichi;Kiyota, Tomomi;Tonganunt, Moltira;Pinkaew, Decha;Liu, Zhihe;Kato, Yoichi;Hutadilok-Towatana, Nongporn;Phongdara, Amornrat;Fujise, Ken
• 通讯作者: Fujise, Ken

Morelloflavone blocks injury-induced neointimal formation by inhibiting vascular smooth muscle cell migration.

• DOI: 10.1016/j.bbagen.2008.09.006
• 发表时间: 2009-01-01
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS
• 影响因子: 3
• 作者: Pinkaew, Decha;Cho, Sung Gook;Hui, David Y.;Wiktorowicz, John E.;Hutadilok-Towatana, Nongporn;Mahabusarakam, Wilawan;Tonganunt, Moltira;Stafford, Lewis J.;Phongdara, Amornrat;Liu, Mingyao;Fujise, Ken
• 通讯作者: Fujise, Ken