ELECTROSTATIC STABILIZATION OF A PROTEIN-DNA COMPLEX

蛋白质-DNA复合体在生物学中非常重要，它控制着染色质结构、DNA复制和蛋白质表达。事实上，转录激活因子和抑制因子的突变或参与DNA复制的蛋白质的突变是癌症的常见原因。更好地了解稳定这些复合体的相互作用将提高我们对DNA结合蛋白突变导致的疾病状态的理解。本提案中描述的项目的目标是确定一套描述静电学在蛋白质-DNA复合体的稳定性和结合亲和力中的作用的一般规则。为了实现这一目标，将采用理论和实验相结合的方法。连续谱静电计算可以提出突变，这些突变将改善Arc抑制子-DNA复合体中的静电互补性，并将在实验中对预测的突变进行测试。此外，还将通过组合诱变技术识别能够改善Arc抑制物与DNA结合的突变，并将进行计算以分析复合体中静电相互作用的变化。

Protein-DNA complexes are extremely important in biology, controlling chromatin structure, DNA replication, and protein expression. In fact, mutations in transcriptional activators and repressors or in proteins involved in DNA replication are common causes of cancer. A greater understanding of the interactions that stabilize these complexes will improve our understanding of the disease states that result from mutations to DNA binding proteins. The goal of the project described in this proposal is to identify a set of general rules describing the role of electrostatics in the stability and binding affinity of protein- DNA complexes. A combination of theoretical and experimental approaches will be used to meet this goal. Continuum electrostatic calculations can suggest mutations that will improve the electrostatic complementarity in the Arc repressor- DNA complex, and the predicted mutations will be tested experimentally. In addition, mutations that improve Arc repressor-DNA binding will also be identified by combinatorial mutagenesis techniques, and calculations will be performed to analyze the changes in the electrostatic interactions in the complex.