BIOLOGICAL ROLES OF THE PROLYL ISOMERASE, PIN1

脯氨酰异构酶 PIN1 的生物学作用

• 批准号: 6514156
• 负责人: ANTHONY R MEANS
• 金额: $ 31.31万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-08-06 至 2004-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6514156
• 关键词: Xenopus oocyte; antisense nucleic acid; cell growth regulation; developmental genetics; enzyme activity; gene deletion mutation; gene targeting; laboratory mouse; laboratory rabbit; leukocyte activation /transformation; lymphocyte; peptidylprolyl isomerase; phenotype; protein binding

Our overall goal is to determine the manner by which the peptidyl-prolyl isomerase, Pin1, acts as a negative regulator of mitosis and evaluate whether this protein may be an attractive target for development of an anti-cancer therapeutic agent. Pin1 was discovered as a protein that interacts with NIMA, a protein Ser/Thr Kinase that is essential for progression from G2 to mitosis in Aspergillus nidulans. Pin1 has been conserved from yeast to man and may be critical for mitotic progression. We cloned Pin1 from Asperigillus and Xenopus, and made antibodies to the protein. We showed that Pin1 will regulate entry into mitosis in Xenopus extracts where it interacts with key components of the p34cdc2 regulatory pathway such as the active forms of the cdc25 protein phosphatase and the plxl protein kinase as well as a number of other mitotic phosphoproteins recognized by the MPM2 antibody. We propose to use the Xenopus Egg/oocyte system to address key issues regarding the prolyl isomerase and protein binding of Pin1 and elucidate critical details of its role(s) as a cell cycle regulator. We will evaluate the tole of Pin1 in the timing of mitotic entry and in the operation of checkpoints governing the G2/M transition as well as examine if Pin1 plays a role in exit from mitosis. In parallel, we will determine if Pin1 is essential for mouse development and focus on its role in development, activation and/or proliferation of T lymphocytes by using the cre/lox system to disrupt the Pin1 gene in mice either globally or specifically in T lymphocytes. We will determine if phenotypic consequences of Pin1 deletion in either vertebrate system requires the prolyl isomerase activity. Completion of these studies will define the role(s) of Pin1 in growth and development, determine its critical cell cycle functions and clarify whether the Pin1 prolyl isomerase is a good candidate for anti-cancer drug development.

我们的总体目标是确定肽基脯氨酰异构酶Pin 1作为有丝分裂负调节因子的方式，并评估这种蛋白质是否可能成为开发抗癌治疗剂的有吸引力的靶点。Pin 1被发现是一种与NIMA相互作用的蛋白质，NIMA是一种蛋白质Ser/Thr激酶，对于构巢曲霉中从G2到有丝分裂的进展至关重要。Pin 1在酵母和人类中是保守的，可能对有丝分裂进程至关重要。我们克隆了Pin 1蛋白，并制备了针对该蛋白的抗体。我们表明，Pin 1将调节进入非洲爪蟾提取物中的有丝分裂，在那里它与p34 cdc 2调节途径的关键组分相互作用，例如cdc 25蛋白磷酸酶和plxl蛋白激酶的活性形式以及MPM 2抗体识别的许多其他有丝分裂磷蛋白。我们建议使用非洲爪蟾卵/卵母细胞系统，以解决关键问题的脯氨酰异构酶和蛋白质结合的Pin 1和阐明其作用的关键细节（S）作为细胞周期调节剂。我们将评估Pin 1在有丝分裂进入的时间和控制G2/M转换的检查点的操作中的托勒，以及检查Pin 1是否在退出有丝分裂中起作用。同时，我们将确定Pin 1是否对小鼠发育至关重要，并通过使用cre/lox系统破坏小鼠中Pin 1基因或在T淋巴细胞中特异性破坏Pin 1基因来关注其在T淋巴细胞发育、活化和/或增殖中的作用。我们将确定Pin 1缺失在任一脊椎动物系统中的表型后果是否需要脯氨酰异构酶活性。这些研究的完成将确定Pin 1在生长和发育中的作用，确定其关键的细胞周期功能，并澄清Pin 1脯氨酰异构酶是否是抗癌药物开发的良好候选者。