Genetic Control of Development

发育的遗传控制

• 批准号: 6921764
• 负责人: JAMES Alfred HOCH
• 金额: $ 7.89万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-05-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6921764
• 关键词: Bacillus subtilis; S adenosylmethionine; affinity chromatography; bacterial genetics; biological signal transduction; developmental genetics; enzyme activity; enzyme substrate; gene expression; gene induction /repression; genetic mapping; genetic transcription; guanine nucleotide binding protein; guanosinetriphosphatases; microarray technology; molecular shape; phosphorylation; protein kinase; regulatory gene; sporogenesis; temperature sensitive mutant; transcription factor; yeast two hybrid system

DESCRIPTION (provided by applicant): Bacteria are highly adaptable organisms capable of growth on countless carbon and nitrogen sources and of occupying an inexhaustible variety of ecological niches including, unfortunately, regions of the human body that are better left bacteria-free. The key to adaptability in bacteria is their capacity to express only those genes for enzymes and pathways that they need for maximal growth in the environment in which they find themselves. One of the major mechanisms of signal recognition leading to specific gene expression is the two-component system and its more complex variant, the phosphorelay. This proposal has the goal of understanding the mechanisms by which both an essential two-component system and the sporulation phosphorelay function, from signal ligand identity to molecular recognition between components and gene activation. Genetic methods will be used to identify signals and proteins activating sporulation sensor kinases. Proteins associating with sensor kinases will be identified. The YycG sensor kinase and YycF response regulator are essential for growth. The genes regulated by this two-component system will be identified by microarray analysis and bioinformatic techniques. Suppression, transposon and multi-copy gene expression techniques will be used to identify the role of the YycF response regulator in gene expression. The role of amino acid side chains in the surface of interaction of response regulators and phosphotransfer domains in recognition specificity will be studied by modifying the Spo0A transcription factor to a sensor kinase substrate. The structure and function of the KinA domains will be studied by domain liberation using mutants bearing specific proteolytic sites in interdomain region. Experiments are proposed to test the hypothesis that the dynamics of the loops making up the active site of Spo0F is an important determinant of sensor kinase specificity and that the dynamics may be influenced by regulating ligands that bind to the response regulator. It is believed that this combination of structural and functional studies will lead to effective anti-bacterial agents either directly or in combination as inhibitors of resistance mechanisms.

描述（由申请人提供）：细菌是能够在无数碳和氮源上生长的高度适应性生物，并且占据了无尽的生态壁ni，包括不幸的是，人体的区域更好，这些区域是更好的剩余细菌。细菌适应能力的关键是它们仅表达这些基因的酶和途径所需的途径，这些基因在他们发现自己的环境中最大程度地生长所需的途径。导致特定基因表达的信号识别的主要机制之一是两个组分系统及其更复杂的变体磷层。该建议的目的是理解从信号配体认同到成分和基因激活之间的分子识别的必不可少的两组分系统和孢子磷函数的机制。遗传方法将用于鉴定激活孢子孢子传感器激酶的信号和蛋白质。将确定与传感器激酶相关的蛋白质。 YYCG传感器激酶和YYCF响应调节剂对于生长至关重要。由微阵列分析和生物信息学技术识别由该两组分系统调节的基因。抑制，转座子和多拷贝基因表达技术将用于识别YYCF响应调节剂在基因表达中的作用。通过将SPO0A转录因子修改为传感器激酶底物，可以研究氨基酸侧链在响应调节剂与磷酸转移域相互作用中的作用。使用域间区域中带有特定蛋白水解位点的突变体，将研究基纳结构域的结构和功能。提出了实验来检验构成SPO0F活性位点的环的动力学的假设是传感器激酶特异性的重要决定因素，并且动力学可能会受到调节与响应调节剂结合的配体的影响。据信，结构和功能研究的这种组合将直接或作为耐药机制抑制剂组合导致有效的抗细菌剂。