C-MYC PHOSPHORYLATION AND GROWTH RELATED KINASES

C-MYC 磷酸化和生长相关激酶

• 批准号: 3458329
• 负责人: BEVERLY BOCKUS
• 金额: $ 9.73万
• 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/3458329
• 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被认为参与了 对细胞增殖的调控。C-myc的异常调控 与细胞的转化有关。尽管广泛 关于c-myc基因水平调控的研究，目前知之甚少 关于c-myc蛋白。这项研究将扩大我们的知识面 关于蛋白质的事。它将强调对后翻译的研究 修饰，特别是磷酸化。生化和 遗传学方法将被用来研究c-myc的作用。 蛋白质在调节细胞增殖中的作用。 基于凝胶的酶和化学切割技术将是 用于分析在许多情况下c-myc的磷酸化模式 生长条件。特定的磷酸化位点将是 经多肽分析、高压电泳法测定 旋转杯中的水解性氨基酸及其序列分析 定序器。将产生针对c-c-的多克隆抗体 MYC/BETS-半乳糖苷酶融合蛋白用于免疫 沉淀天然蛋白质和变性蛋白质。 C-myc的位点特异性突变体将使用 寡核苷酸导向的突变。这些突变体将会是 进行了功能和生物学特性测试。 这项研究还将检查其他癌基因的影响，如 作为多瘤T抗原对c-myc的磷酸化。多发瘤 不能刺激宿主细胞DNA合成的突变体 将产生并测试它们对细胞生长的影响 相关的激酶和c-myc的磷酸化。