BIOLOGICAL ROLES OF THE PROLYL ISOMERASE, PIN1

脯氨酰异构酶 PIN1 的生物学作用

• 批准号: 6377435
• 负责人: ANTHONY R MEANS
• 金额: $ 30.51万
• 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/6377435
• 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 Pin1 作为有丝分裂负调节因子的作用方式，并评估该蛋白是否可能成为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 是一种丝氨酸/苏氨酸激酶蛋白，对于构巢曲霉从 G2 期进展到有丝分裂至关重要。 Pin1 从酵母到人类都是保守的，可能对有丝分裂进程至关重要。我们从曲霉属和非洲爪蟾属中克隆了 Pin1，并制备了该蛋白的抗体。我们表明，Pin1 将调节非洲爪蟾提取物中进入有丝分裂的过程，其中它与 p34cdc2 调节途径的关键成分相互作用，例如 cdc25 蛋白磷酸酶和 plxl 蛋白激酶的活性形式，以及许多其他被有丝分裂蛋白识别的有丝分裂磷蛋白。 MPM2 抗体。我们建议使用非洲爪蟾卵/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作用的关键细节。我们将评估 Pin1 在有丝分裂进入时间和控制 G2/M 转换的检查点操作中的作用，并检查 Pin1 是否在有丝分裂退出中发挥作用。同时，我们将确定 Pin1 是否对小鼠发育至关重要，并通过使用 cre/lox 系统全局或特异地破坏小鼠 T 淋巴细胞中的 Pin1 基因，重点关注其在 T 淋巴细胞发育、激活和/或增殖中的作用。我们将确定任一脊椎动物系统中 Pin1 缺失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 Pin1 在生长和发育中的作用，确定其关键的细胞周期功能，并阐明 Pin1 脯氨酰异构酶是否是抗癌药物开发的良好候选者。