GENETIC AND BIOCHEMICAL ANALYSIS OF CELL BEHAVIOR

细胞行为的遗传和生化分析

• 批准号: 3916342
• 负责人: M M GOTTESMAN
• 金额: --
• 依托单位: DIVISION OF CANCER BIOLOGY AND DIAGNOSIS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3916342
• 关键词: Rous sarcoma virus; cell growth regulation; cyclic AMP; cytogenetics; fibroblasts; gel electrophoresis; gene expression; genetic library; genetic manipulation; genetic promoter element; interferons; molecular cloning; molecular oncology; morphology; mutant; neoplasm /cancer genetics; nucleic acid sequence; protein kinase; protein kinase A; protein structure; tissue /cell culture; transfection; transposon /insertion element

We are utilizing the Chinese hamster ovary (CHO) fibroblast to study the genetics and biochemistry of some aspects of the behavior of cultured cells. Our work has emphasized morphology and its relationship to growth control, and the manner in which cyclic AMP regulates cell growth and gene expression. The mechanism of cAMP action on CHO cells has been studied by isolating CHO mutants resistant to growth inhibition by cAMP. These mutants have defective cAMP dependent protein kinases with either altered regulatory (RI) or catalytic (C) subunits. DNA from cells carrying dominant cAMP-resistant defects can be used to transfer the cAMP- resistance phenotype to sensitive cells. Cosmid clones carrying the CHO RI subunit from cAMP resistant cells have been isolated. Sequence analysis indicates that one of these clones, pcosRI has the coding sequence for the CHO RI subunit. The exon-intron structure of the RI gene is complex; the region encoding the cAMP- binding site at the 3' end of the gene is composed of several different exons. The regulation of expression of cAMP-responsive genes is being studied using wild type, cAMP dependent protein kinase mutants, and transferent CHO cell lines expressing the mutant kinase phenotype. We have demonstrated that for the rat tyrosine amino transferase (TAT) gene promoter an intact cAMP dependent protein kinase system is necessary for the regulation of cAMP responsive gene expression.

我们正在利用中国仓鼠卵巢（CHO）成纤维细胞， 研究行为的某些方面的遗传学和生物化学， 培养的细胞。 我们的工作强调了形态学及其 与生长控制的关系，以及环磷酸腺苷 调节细胞生长和基因表达。 cAMP的作用机制 通过分离CHO突变体研究了对CHO细胞的作用 对cAMP的生长抑制有抗性。 这些变种人 cAMP依赖性蛋白激酶缺陷， 调节（RI）或催化（C）亚基。DNA来自携带 显性cAMP抗性缺陷可用于转移cAMP- 对敏感细胞的抗性表型。 宇宙克隆携带 已从cAMP抗性细胞中分离出CHO RI亚基。 序列分析表明，这些克隆之一，pcosRI具有 CHO RI亚基的编码序列。 外显子-内含子 RI基因的结构是复杂的;编码cAMP的区域- 基因3'端的结合位点由几个 不同的外显子 cAMP反应性的表达调控 基因正在使用野生型cAMP依赖性蛋白进行研究 激酶突变体和表达该激酶突变体的转染CHO细胞系。 突变激酶表型。 我们已经证明，对于老鼠来说， 酪氨酸氨基转移酶（达特）基因启动子和完整的cAMP 依赖性蛋白激酶系统是调节 cAMP应答基因表达。