DNA SYNTHESIS: REGULATION IN NORMAL AND CANCER CELLS

DNA 合成：正常细胞和癌细胞中的调节

• 批准号: 3164118
• 负责人: EARL F BARIL
• 金额: $ 33.12万
• 依托单位: WORCESTER FOUNDATION FOR BIOMEDICAL RES
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-04-01 至 1991-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3164118
• 关键词: DNA directed DNA polymerase; HeLa cells; antigens; binding proteins; cell cycle; cell growth regulation; chromatography; conformation; electron microscopy; enzyme complex; enzyme mechanism; enzyme structure; gel electrophoresis; laboratory rabbit; neoplasm /cancer genetics; neoplastic cell; nucleoside triphosphate; protein reconstitution; simian virus 40; tissue /cell culture

The objective of the research is to define possible controls for DNA biosynthesis during normal and abnormal growth. Most of the DNA polymerase Alpha activity from HeLa cells in S-phase has been isolated as a Mr 640,000 multiprotein form that is designated DNA polymerase Alpha2. The enzyme has been purified to electrophoretic homogeneity and resolved into its components that include; a Mr 180,000 polymerase Alpha subunit, a Mr 70,000 DNA primase, a Mr 69,000 3'---greater than 5' exonuclease, Mr 96,000 C1 and Mr 52,000 C2 primer-recognition proteins and Mr 92,000 Ap A binding protein. The interaction of DNA polymerase Alpha2 and other required proteins for the efficient and specific initiation of RNA primer synthesis will be defined. The conditions and protein requirements in addition to polymerase Alpha2 that are required for elongation of DNA chains with catalytic efficiency, processivity and fidelity will be ascertained. The enzymic and nonenzymic proteins in addition to DNA polymerase Alpha2 that are required for the replicationof SV40-DNA in vitro in the presence of T-antigen will be studied.

这项研究的目的是确定可能的控制DNA 在正常和异常生长期间的生物合成。 大多数DNA聚合酶 来自S期HeLa细胞的α活性已被分离为Mr 640，000 一种多蛋白形式，被命名为DNA聚合酶α 2。 所述酶具有 纯化至电泳均一，并拆分成其 包括：一个Mr 180，000聚合酶α亚基，一个Mr 70，000聚合酶α亚基， DNA引发酶，一种Mr 69，000的3 '-大于5'的核酸外切酶，Mr 96，000的C1和 Mr 52，000 C2引物识别蛋白和Mr 92，000 Ap A结合 蛋白 DNA聚合酶α 2和其他必需的 用于RNA引物合成的有效和特异性起始的蛋白质 将被定义。 条件和蛋白质的要求，除了 聚合酶α 2是延长DNA链所需的， 将确定催化效率、持续合成能力和保真度。 的 除了DNA聚合酶α 2之外， 是体外SV 40-DNA复制所必需的 将研究T抗原。