Polymerization and Activation of Bacterial RecA Proteins

细菌 RecA 蛋白的聚合和激活

• 批准号: 6701883
• 负责人: FREDERICK W WHIPPLE
• 金额: $ 20.1万
• 依托单位: CALIFORNIA STATE UNIVERSITY FULLERTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-01-01 至 2007-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6701883
• 关键词: bacterial genetics; bacterial proteins; enzyme structure; gene mutation; genetic manipulation; genetic recombination; polymerization; protein protein interaction; protein sequence; protein structure function; recombinase; suppressor mutations

DESCRIPTION (provided by applicant): Bacterial genetic techniques will be used to investigate the mechanism by which molecules of the RecA protein of the bacterium Escherichia coli polymerize around DNA to form an active nucleoprotein complex. Proper formation of this complex is essential to the processes of DNA repair and DNA recombination, both of which are necessary for survival. RecA is closely related to similar proteins found in cells of higher organisms, including humans. This work is relevant to human health because defects in DNA recombination and repair in humans usually lead to cancer or severe genetic disorders. A deeper understanding of these processes may help in the development of treatments and diagnostic techniques for these specific illnesses. The genetic technique of mutant-suppressor pair analysis will be applied to this problem. This involves the targeting of a mutation to one member of a pair of interacting proteins, and then seeking second mutations in the other member of the pair that reverse the effect of the first mutation. Mutant pairs such as this are very useful in mapping individual amino acid contact points between interacting proteins. To implement this strategy, an altered form of the RecA polymer will first be generated. This will done by obtaining pair of recA mutants that are each unable to polymerize by themselves, but are able to interact with each other to form fully functional polymers. These polymers will be characterized by alternating interface specificities.

描述(申请人提供)：细菌遗传技术将被用来研究细菌的RecA蛋白分子围绕DNA聚合形成活性核蛋白复合体的机制。这种复合体的正确形成对DNA修复和DNA重组过程至关重要，而DNA修复和DNA重组是生存所必需的。RecA与高等生物细胞中发现的类似蛋白质密切相关，包括人类。这项工作与人类健康相关，因为人类DNA重组和修复的缺陷通常会导致癌症或严重的遗传疾病。对这些过程的深入了解可能有助于开发针对这些特定疾病的治疗和诊断技术。 突变-抑制子对分析的遗传技术将应用于这一问题。这涉及到将突变定位于一对相互作用的蛋白质中的一个成员，然后在该对中的另一个成员中寻找第二个突变，以逆转第一个突变的效果。像这样的突变对在相互作用的蛋白质之间定位单个氨基酸接触点是非常有用的。为了实施这一策略，首先将产生一种改变形式的RecA聚合物。这将通过获得一对recA突变体来实现，这些突变体都不能自行聚合，但能够相互作用形成功能齐全的聚合物。这些聚合物的特征是具有交替的界面特性。