C-MYC PHOSPHORYLATION AND GROWTH RELATED KINASES

C-MYC 磷酸化和生长相关激酶

• 批准号: 3458328
• 负责人: BEVERLY BOCKUS
• 金额: $ 9.98万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-08-01 至 1993-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3458328
• 关键词: DNA; DNA binding protein; affinity labeling; antibody; antigens; autoradiography; cell cycle; cell growth regulation; cell transformation; cytogenetics; electrofocusing; enzyme linked immunosorbent assay; gene interaction; genetic mapping; genetic translation; high voltage electrophoresis; immunological substance; immunoprecipitation; laboratory mouse; laboratory rabbit; molecular genetics; molecular site; mutagen testing; nucleic acid hybridization; phosphorylation; protein kinase; protein sequence; protooncogene; radiotracer; tissue /cell culture

The nuclear proto-oncogene c-myc is thought to be involved in regulation of cell proliferation. Abnormal regulation of c-myc has been implicated in transformation of cells. Despite extensive work on the regulation of c-myc mRNA levels, little is known about the c-myc protein. This study will expand our knowledge about the protein. It will emphasize studies on post-translation modifications, particularly phosphorylation. Biochemical and genetic approaches will be used to examine the role of c-myc protein in the regulation of cellular proliferation. Gel-based enzymatic and chemical cleavage techniques will be used to analyze c-myc phosphorylation patterns under a number of growth condition. Specific phosphorylation sites will be determined by peptide analysis, high voltage electrophoresis of hydrolyzed amino acids and sequence analysis in a spinning cup sequenator. Polyclonal antibodies will be generated against a c- myc/bets-galactosidase fusion protein and used to immuno- precipitate native and denatured protein. Site specific mutants of c-myc will be constructed using oligonucleotide directed mutagenesis. These mutants will be tested for functional and biological properties. This study will also examine the effects of other oncogenes such as the polyoma T antigens on c-myc phosphorylation. Polyoma mutants defective for the stimulation of host cell DNA synthesis will be generated and tested for their effects on cellular growth related kinases and c-myc phosphorylation.

核原癌基因 c-myc 被认为参与 细胞增殖的调节。 c-myc 调节异常 与细胞转化有关。 尽管广泛 有关 c-myc mRNA 水平调节的工作，目前知之甚少 关于c-myc蛋白。 这项研究将扩展我们的知识 关于蛋白质。 它将强调翻译后研究 修饰，特别是磷酸化。 生化和 遗传方法将用于检查 c-myc 的作用 调节细胞增殖的蛋白质。 基于凝胶的酶促和化学裂解技术将 用于分析多种条件下的 c-myc 磷酸化模式 生长状况。 具体的磷酸化位点是 通过肽分析、高压电泳测定 旋转杯中的水解氨基酸和序列分析 序列器。 将产生针对 c-的多克隆抗体 myc/bets-半乳糖苷酶融合蛋白并用于免疫- 沉淀天然和变性蛋白质。 c-myc 的位点特异性突变体将使用构建 寡核苷酸定向诱变。 这些突变体将是 测试了功能和生物特性。 这项研究还将检查其他致癌基因的影响，例如 作为 c-myc 磷酸化的多瘤 T 抗原。 多马 刺激宿主细胞 DNA 合成有缺陷的突变体 将生成并测试它们对细胞生长的影响 相关激酶和 c-myc 磷酸化。