PATHWAY LEADS TO APOPTOSIS IN SRC-TRANSFORMED CELLS

SRC 转化细胞中导致凋亡的途径

• 批准号: 2009247
• 负责人: Hwa-Chain Robert Wang
• 金额: $ 9.95万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-04-01 至 1997-10-10
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2009247
• 关键词: SDS polyacrylamide gel electrophoresis; affinity chromatography; animal tissue; antisense nucleic acid; apoptosis; biological signal transduction; cell transformation; cellular oncology; complementary DNA; enzyme activity; immunofluorescence technique; immunoprecipitation; molecular cloning; peptide chemical synthesis; phosphorylation; polymerase chain reaction; protein kinase; protein structure function; western blottings

Long terms goals of my research are to understand intracellular signaling controls altered upon cellular transformation and to restore the control to reverse the progress of cancers by suppressing the transforming features or driving transformed cells to apoptosis. Ultimate goal of this project is to identify signaling pathway (s) which link (s) cellular transformation to apoptosis. I have detected a novel serine/threonine protein kinase, p63 SAMK (src activated MBP kinase) whose activation may be involved in the process of apoptosis in normal and transformed cultures of avian and mammalian cells. Recently, we isolated a cDNA for a human protein kinase Krs-1 (kinase responsive to stress) who catalytic domain shares extensive homology with Ste20p family. Although Krs-1 shares many properties with p63 SAMK, discrepancy in the antigenicity and the analyzed peptide sequences raises a question of the homology between Krs-1 and p63 SAMK. In this project, I propose the following studies: 1. TO clarify the identify of chick p63 SAMK and the relationship with Krs-1 by isolation of the chick Krs-1 homologue cDNS and further study of the antigenic similarity between p63 SAMK and Krs-1. 2. To determine that nature of the activator of Krs-1/p63 SAMK in signaling cascade by biochemical study and chromatography. 3. To study the biological role of Krs-1/p63 SAMK in cellular transformation and apoptosis by expression of the gene products including the wild-type and mutant proteins and antisense messages in src-transformed or normal cells. Success of this project will advance our understanding of a particular signaling pathway linking cellular transformation to apoptosis-related cellular events.

我研究的长期目标是了解细胞内信号传导 细胞转化后控制发生改变并恢复控制 通过抑制转化特征来逆转癌症的进展 或驱动转化细胞凋亡。 该项目的最终目标 是确定连接细胞转化的信号通路 至细胞凋亡。 我检测到一种新的丝氨酸/苏氨酸蛋白激酶，p63 SAMK（src 激活的MBP激酶）其激活可能参与以下过程 鸟类和哺乳动物细胞的正常和转化培养物中的细胞凋亡。 最近，我们分离出了人类蛋白激酶Krs-1（kinase 对压力敏感）谁的催化结构域具有广泛的同源性 Ste20p家族。 尽管 Krs-1 与 p63 SAMK 共享许多特性， 抗原性和分析的肽序列之间存在差异 Krs-1 和 p63 SAMK 之间的同源性问题。 在这个项目中， 我建议进行以下研究： 1. 明确chick p63的鉴定 SAMK 以及通过分离雏鸡 Krs-1 与 Krs-1 的关系 同源cDNS及p63之间抗原相似性的进一步研究 SAMK 和 Krs-1。 2. 确定 Krs-1/p63 SAMK 激活剂的性质 通过生化研究和色谱法进行信号级联反应。 3. 研究Krs-1/p63 SAMK在细胞中的生物学作用 通过表达基因产物进行转化和凋亡，包括 src 转化中的野生型和突变蛋白以及反义信息 或正常细胞。 该项目的成功将加深我们对特定领域的理解 连接细胞转化与凋亡相关的信号通路 细胞事件。