Polymerization and Activation of Bacterial RecA Proteins

描述（由申请人提供）：细菌遗传技术将用于研究大肠杆菌RecA蛋白分子在DNA周围聚合形成活性核蛋白复合物的机制。这种复合体的正确形成对DNA修复和DNA重组的过程至关重要，这两者都是生存所必需的。RecA与包括人类在内的高等生物细胞中发现的类似蛋白质密切相关。这项工作与人类健康有关，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。

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.