DNA MODIFICATION RESTRICTION IN PRO AND EUKARYOTES

原核生物和真核生物中的 DNA 修饰限制

• 批准号: 3276786
• 负责人: STANLEY M HATTMAN
• 金额: $ 30.31万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 1980
• 资助国家: 美国
• 起止时间: 1980-09-01 至 1994-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3276786
• 关键词: 5 methylcytosine; DNA binding protein; DNA methylation; DNA replication; Drosophilidae; Escherichia coli; azacitidine; bacterial DNA; bacterial genetics; bacterial proteins; bacteriophage T4; biochemical evolution; chickens; chromatin; density gradient ultracentrifugation; endonuclease; eukaryote; gene expression; gene mutation; genetic manipulation; genetic mapping; genetic promoter element; genetic transcription; genetic translation; high performance liquid chromatography; host organism interaction; laboratory rat; lac operon; mass spectrometry; methyltransferase; molecular cloning; mutagens; nucleic acid sequence; nucleic acid structure; plasmids; prokaryote; protein biosynthesis; protein engineering; protein sequence; proteins; radiotracer; transcription factor; yeasts

Our long term objective is to gain a greater understanding of postreplicative DNA modifications, with respect to their nature, mechanism and specificity of occurrence and their role in gene expression. We are primarily interested in normally occurring modifications controlled by cells or viruses (viz. enzymatic processes, rather than chemically-induced modification) since these are more likely to be of biological importance in the growth and development of the organism. Specifically, we are focusing on the DNA-adenine methyltransferase (Dam) of Escherichia coli and phages T2 and T4. We hope to elucidate the domains involved in sequence/substrate recognition and, eventually, the precise mechanism underlying polypeptide-DNA interaction. By site-directed mutagenesis and in vitro recombination, we will attempt to generate enzymes with new sequence specificities. In a separate system (involving phage Mu), we are in the process of defining the various regulatory process by which expression of the DNA modification gene (mom) is controlled. The detailed mechanisms of action of the Mu C (transcription factor), Mu Com (translation factor) and host E. coli Dam methylase (transcription factor) are under investigation.

我们的长期目标是更好地了解 复制后 DNA 修饰，就其性质而言， 发生的机制和特异性及其在基因中的作用 表达。 我们主要对正常发生的情况感兴趣 由细胞或病毒控制的修饰（即酶促 过程，而不是化学诱导的修饰），因为这些 在生长过程中更有可能具有生物学重要性 有机体的发育。 具体而言，我们重点关注 大肠杆菌和噬菌体的 DNA-腺嘌呤甲基转移酶 (Dam) T2 和 T4。 我们希望阐明所涉及的领域 序列/底物识别，并最终精确 多肽-DNA相互作用的机制。 通过站点定向 诱变和体外重组，我们将尝试产生 具有新序列特异性的酶。 在一个单独的系统（涉及噬菌体 Mu）中，我们正在处理 定义各种监管程序，通过该程序表达 DNA修饰基因（mom）受到控制。 详细的 Mu C（转录因子）、Mu Com 的作用机制 （翻译因子）和宿主大肠杆菌 Dam 甲基化酶（转录因子） 因素）正在调查中。