GENETIC APPROACHES TO PROTEIN-NUCLEIC ACID INTERACTIONS

蛋白质-核酸相互作用的遗传学方法

• 批准号: 2174118
• 负责人: PAUL R SCHIMMEL
• 金额: $ 28.49万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-03-01 至 1997-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2174118
• 关键词: Escherichia coli; Saccharomyces cerevisiae; X ray crystallography; acylation; alanine; aminoacid tRNA ligase; aminoacyl tRNA; biochemical evolution; chemical fingerprinting; crosslink; enzyme induction /repression; enzyme mechanism; enzyme structure; genetic manipulation; glutamine; hybrid enzyme; isoleucine; methionine; mutant; nucleic acid sequence; protein engineering; site directed mutagenesis; temperature sensitive mutant; transfer RNA

Aminoacyl transfer RNA synthetases arose early in evolution and established the rules of the genetic code through their specific aminoacylations of transfer RNAs. the enzymes are organized into two classes of ten enzymes each. These two classes appear to have no historical relationship to each other. Experiments are proposed to investigate and manipulate the tRNA specificity of class I and class II enzymes. These synthetases include E. coli alanine, glutamine, isoleucine and methionine tRNA synthetases, and S. cerevisiae glutamine tRNA synthetase. Recent experiments have shown that the amino acid acceptor helix is a major determinant of the identity of at least some tRNAs. Functional synthetase-tRNA acceptor helix contacts will be identified and manipulated through analysis of mutant enzymes which are selected for specific alterations in their acceptor helix interactions. A minimalist aminoacyl tRNA synthetase will also be designed and subjected to mutagenesis and selection for functional activity. Additional efforts will be directed at manipulations of tRNA recognition specificity by introducing the fewest possible amino acid or nucleotide substitutions, in some cases taking advantage of recent x-ray crystallographic structural information.

氨基酰基转移RNA合成酶在进化早期出现 通过其具体建立了遗传密码的规则 转移RNA的氨基酰胺。 将酶组织成两个 每个酶的类别。 这两个课程似乎没有 彼此的历史关系。 提出了实验 调查和操纵I类和班级的tRNA特异性 II酶。 这些合成酶包括大肠杆菌丙氨酸，谷氨酰胺， 异亮氨酸和蛋氨酸TRNA合成酶，以及酿酒酵母谷氨酰胺 tRNA合成酶。 最近的实验表明氨基酸 受体螺旋是至少某些身份的主要决定因素 trnas。 功能性合成酶-TRNA受体螺旋触点将是 通过分析突变酶来识别和操纵 选择用于其受体螺旋相互作用的特定更改。 简约的氨基酰基TRNA合成酶也将设计和 受到诱变和功能活性的选择。 额外的努力将致力于操纵tRNA识别 通过引入最少可能的氨基酸或核苷酸来特异性 替换，在某些情况下利用最近的X射线 晶体学结构信息。