Protein Architecture and Remodeling in DNA Transposition and DNA Protection

高级蛋白质-DNA复合物协调和催化许多生命最核心的过程。 基因重组和DNA保护，维持基因组完整性的关键过程 对于后代来说，尤其是在蛋白质-DNA超结构的使用方面。其中的一个关键特征是 复合物的另一个特点是它们通常非常稳定，这样ATP依赖性蛋白质解折叠酶 并且可能需要蛋白酶来重塑、分解、破坏或回收组分蛋白质。的 该项目的重点是了解蛋白质复合物是如何被识别为重塑或破坏， Clp/Hsp 100 ATP酶家族的蛋白质。Clp/Hsp 100蛋白是AAA+酶的亚家族， 使用ATP水解在其底物上进行机械功。第一个具体目标是了解 蛋白质解折叠酶ClpX如何识别蛋白质-DNA复合物， 噬菌体Mu的转座。阐明转座酶内肽信号的实验 的认可提出。此外，我们将测试一个模型，在该模型中， 复合物将ClpX的解折叠活性引导到一个特定的转座酶亚基，从而产生一个特异性的转座酶亚基。 新的-不太稳定-复杂的独特的架构。第二个目标是阐明DNA- 保护蛋白Dps被ClpX识别，以及这种识别如何适应不断变化的环境 条件将研究肽信号、衔接蛋白和DNA在识别Dps中的作用。 最后，通过蛋白质组学实验设计，实现了对ATP依赖蛋白作用的全局观察 提出了解折叠酶/蛋白酶。这些实验将提供一个蛋白质组范围内的观点， 特异性衔接蛋白和肽结合结构域在AAA+酶的底物选择中的作用。蜂窝 保护基因组免受损伤并促进受损DNA的忠实修复的机制是 对健康和生存至关重要，正如许多癌症综合征所证明的那样， 这些细胞过程的遗传破坏。AAA+引起的蛋白质重塑和破坏 酶是细胞用于解释和解释细胞的策略的一个关键但知之甚少的方面， 保护他们的基因组

Higher-order protein-DNA complexes both orchestrate and catalyze many of life's most central processes. Genetic recombination and DMAprotection, processes essential for maintaining the integrity of the genome for future generations, are especially rich in their use of protein-DNA superstructures. A key feature of these complexes is that they are often exceedingly stable, such that ATP-dependent protein-unfolding enzymes and proteases may be required to remodel, dismantle, destroy or recycle the component proteins. The focus of this project is to understand how protein complexes are recognized for remodeling or destruction by proteins of the Clp/Hsp100 ATPase family. Clp/Hsp100 proteins are a subfamily of the AAA+ enzymes that use ATP-hydrolysis to perform mechanical work on their substrates. The first specific goal is to understand how the protein-unfolding enzyme CIpX recognizes the protein-DNA complex that promotes DNA transposition of phage Mu. Experiments to elucidate the peptide signals within the transposase responsible for recognition are proposed. Furthermore, we will test a model in which asymmetric features of the complex guide the unfolding activity of CIpX to one specific transposase subunit, and thereby generate a new -less stable-complex with a unique architecture. The second goal is to elucidate how the DNA- protection protein Dps is recognized by CIpX and how this recognition is tuned to changing environmental conditions. The role of peptide signals, adaptor proteins and DNA in recognition of Dps will be investigated. Finally, proteomic experiments designed to achieve a global view of the role of ATP-dependent protein unfoldases/proteases are proposed. These experiments will give a proteome-wide view of the roles of specific adaptor proteins and peptide binding domains in substrate choice by AAA+ enzymes. Cellular mechanisms that protect the genome from damage, and promote the faithful repair of damaged DNA are critical to health and survival, as dramatically demonstrated by the numerous cancer syndromes associated with the genetic disruption of these cellular processes. Protein remodeling and destruction byAAA+ enzymes is a critical yet poorly understood aspect of the strategies used by cells to both interpret and protect their genomes.