P33ING1 IN CELLULAR SENESCENCE AND ORGANISIMAL AGING

P33ING1 在细胞衰老和组织衰老中的作用

• 批准号: 2826840
• 负责人: JUNYING YUAN
• 金额: $ 27.98万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2826840
• 关键词: RNase protection assay; aging; apoptosis; binding sites; cell growth regulation; cell proliferation; cell senescence; gene expression; gene targeting; genetic regulation; genetically modified animals; laboratory mouse; p53 gene /protein; phosphatidylinositol 3 kinase; phosphatidylinositols; point mutation; protein localization; protein structure function; site directed mutagenesis; superoxide dismutase; transcription factor; tumor suppressor genes

The long-term objective of this proposal is to illustrate the mechanisms and functions of tumor suppressor gene p33ING1 in regulation of cellular senescence and organismal aging. This work will be a collaboration with the laboratories of Ruvkun, Avruch and Alexander-Bridges. The work from Ruvkun's lab showed that the longevity of C. elegans is at least partly regulated by a pathway involving daf-2, an insulin receptor-like gene, and age-1, a homologue of the mammalian phosphatidylinositol-3-OH kinase (PI3 kinase) catalytic subunit. We hypothesize that the aging of mammalian animals is regulated by a similar pathway involving homologues the genes in the C. elegans age-1 pathway. In a screen designed to identify phosphatidylinositol (3,4,5)- triphosphate (PtdIns(3,4,5)P3 binding proteins, we found that PtdIns(3,4,5)P3 can bind specifically to p33ING1, a mammalian tumor suppressor gene product. p33ING1 has been shown to interact physically with p53 in mediating cellular growth control and senescence. We propose that regulation of p33ING1 by PI3 kinase plays an important role in regulating cell proliferation and senescence. p33ING1 may be a critical missing link between the growth factor signal transduction pathways mediated through PI3 kinase and cell proliferation and senescence regulated by tumor suppressor genes. Specific Aim 1 is to determine the functional interaction between PI3 kinase and p33ING1 in mediating cellular growth control and senescence. Our hypothesis is that the activation of Pi3 kinase suppresses p33ING1 activity and its ability to induce cellular senescence and apoptosis. Specific Aim 2 is to characterize the p53/p33ING1 complex and the role of PtdIns(3,4,5)P3 binding in complex formation. We would like to determine the proteins associated with p53/p33ING1 complex and the role of PI3 kinase in the interaction. Specific Aim 3 is to determine the downstream events regulated by p33ING1. We would like to determine whether p33ING1 acts upstream or in a parallel pathway of the mammalian homologues of DAF-16. Specific Aims 4 is to determine the in vivo functions of p33ING1 by generation and characterization p33ING1 mutant mice generated by gene-targeting. We would like to generate mutant mice expressing a null allele or a constitutively active mutant allele of p33ING1. These works will illustrate the role of p33ING1 in mediating cellular senescence in culture and organismal aging in vivo.

该提案的长期目标是阐明抑癌基因p33ING1在调节细胞衰老和机体衰老中的机制和功能。 这项工作将与 Ruvkun、Avruch 和 Alexander-Bridges 的实验室合作进行。 Ruvkun 实验室的工作表明，线虫的寿命至少部分受到涉及 daf-2（一种胰岛素受体样基因）和 Age-1（哺乳动物磷脂酰肌醇-3-OH 激酶同源物）的通路的调节。 PI3 激酶）催化亚基。 我们假设哺乳动物的衰老是由类似的途径调节的，该途径涉及线虫age-1途径中基因的同源物。在旨在鉴定磷脂酰肌醇 (3,4,5)-三磷酸（PtdIns(3,4,5)P3 结合蛋白）的筛选中，我们发现 PtdIns(3,4,5)P3 可以与哺乳动物肿瘤 p33ING1 特异性结合p33ING1 已被证明与 p53 物理相互作用，介导细胞生长控制和衰老。 p33ING1 在调节细胞增殖和衰老中发挥重要作用，可能是通过 PI3 激酶介导的生长因子信号转导途径与肿瘤抑制基因调节的细胞增殖和衰老之间的关键缺失环节。 PI3 激酶和 p33ING1 介导细胞生长控制和衰老 我们的假设是 Pi3 激酶的激活抑制 p33ING1 活性及其诱导细胞衰老和凋亡的能力。具体目标 2 是表征 p53/p33ING1 复合物以及 PtdIns(3,4,5)P3 结合在复合物形成中的作用。 我们想要确定与 p53/p33ING1 复合物相关的蛋白质以及 PI3 激酶在相互作用中的作用。 具体目标 3 是确定 p33ING1 调节的下游事件。我们想要确定 p33ING1 是否在 DAF-16 哺乳动物同源物的上游或平行途径中发挥作用。 具体目标 4 是通过基因靶向产生的 p33ING1 突变小鼠的生成和表征来确定 p33ING1 的体内功能。 我们希望产生表达 p33ING1 无效等位基因或组成型活性突变等位基因的突变小鼠。 这些工作将说明 p33ING1 在介导培养细胞衰老和体内有机体衰老中的作用。