BIOLOGICAL ROLES OF THE PROLYL ISOMERASE, PIN1

脯氨酰异构酶 PIN1 的生物学作用

• 批准号: 6895404
• 负责人: ANTHONY R MEANS
• 金额: $ 5.22万
• 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 2005-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6895404
• 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 - 丙烯基异构酶PIN1充当有丝分裂的负调节剂的方式，并评估该蛋白是否可能是抗癌治疗剂开发的有吸引力的靶标。 PIN1被发现是一种与NIMA相互作用的蛋白质，NIMA是一种蛋白Ser/Thr激酶，对于从G2到曲曲霉中的有丝分裂至关重要。 PIN1从酵母到人都保存，可能对有丝分裂进展至关重要。我们将PIN1从曲折和爪蟾中克隆，并制成了蛋白质的抗体。我们表明，PIN1将调节Xenopus提取物与p34CDC2调节途径的关键成分相互作用，例如CDC25蛋白磷酸酶的活性形式和PLXL蛋白激酶的活性形式以及许多其他有丝分裂磷酸蛋白，由许多其他有丝分裂磷酸蛋白认识到。 MPM2抗体。我们建议使用爪蟾鸡蛋/卵母细胞系统来解决有关PIN1的脯氨酰异构酶和蛋白质结合的关键问题，并阐明其作为细胞周期调节剂的关键细节。我们将在有丝分裂进入的时间和控制G2/M转变的检查点的操作中评估PIN1的刺激，并检查PIN1是否在有丝分裂的退出中起作用。同时，我们将通过使用CRE/LOX系统来破坏全球或特定于T淋巴细胞中小鼠中的PIN1基因来破坏T PIN1是否对于小鼠发育至关重。我们将确定在任一个脊椎动物系统中PIN1缺失的表型后果是否需要丙酰同异构酶活性。这些研究的完成将定义PIN1在生长和发育中的作用，确定其关键细胞周期功能，并阐明PIN1脯氨酸异构酶是否是抗癌药物开发的良好候选者。