Physiological control of Mu-dependent DNA arrangements

Mu依赖性DNA排列的生理控制

• 批准号: 9220089
• 负责人: James Shapiro
• 金额: $ 28.5万
• 依托单位: University of Chicago
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-02-15 至 1997-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9220089&HistoricalAwards=false
• 关键词: Physiological; control; Mu; dependent; DNA

Formation of araB-lacZ hybrid coding sequences normally utilizes the DNA rearrangement activities of transposable bacteriophage Mu. Fusion events are physiologically specific: they do not occur during normal growth but are found at high rates following starvation. This system is a prime example of inducible genetic changes now called "directed mutation." We have developed a detailed molecular explanation of the complex series of biochemical events that produce ara-lac fusions. The cAMP-CRP catabolite sensory system and the CLP heat-shock protease play essential roles in Mu activation during starvation. PCR technology has been validated for the detection of rare fusion events, and there is evidence for no-fusion rearrangements after stress. Thus, the ara- lac fusion system is one of the best-characterized physiologically regulated mutational systems. Genetic studies and molecular experiments are outlined here to define the roles of the cAMP-CRP and CLP systems in Mu activation, to identify alternative biochemical pathways to fusion formation, and to clarify the nature of Mu-dependent DNA rearrangements following stress. %%% The results of this research will provide definitive answers to several issues in the directed mutation controversy and identify components of the signal transduction network connecting cellular physiology and DNA biochemistry.

araB-lacZ杂合编码序列的形成通常利用 转座噬菌体Mu DNA重排活性。 融合事件是生理学上特异性的：它们不会发生 在正常生长期间，但在以下情况下， 饥饿 这个系统是诱导遗传学的一个很好的例子 现在被称为“定向突变”。“我们开发了一种 详细的分子解释的复杂系列的生化 产生ara-lac融合的事件。 cAMP-CRP分解代谢物 感觉系统和CLP热激蛋白酶起着重要作用 在饥饿时的Mu激活。 PCR技术已经 经验证可用于检测罕见融合事件， 应力后无融合重排的证据。 因此，阿拉- 紫胶融合系统是最具特征的生理学 调节突变系统。 遗传学研究和分子生物学 这里概述了实验来定义cAMP-CRP的作用 和CLP系统在Mu激活，以确定替代 融合形成的生物化学途径，并澄清 依赖于mu的DNA重排的过程 %%% 这项研究的结果将提供明确的答案， 定向突变争议与鉴别中的几个问题 信号转导网络的组成部分， 生理学和DNA生物化学。