Protein Architecture and Remodeling in DNA Transposition and DNA Protection

DNA 转座和 DNA 保护中的蛋白质结构和重塑

• 批准号: 7224155
• 负责人: TANIA A BAKER
• 金额: $ 28.84万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-05-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7224155
• 关键词: ATP Hydrolysis; ATP phosphohydrolase; Adaptor Signaling Protein; Affinity; Architecture; Area; Bacteriophage mu; Binding; Binding Proteins; Biochemical Genetics; Cell physiology; Cells; Complex; Condition; DNA; DNA Binding; DNA Damage; DNA Repair; Disruption; Endopeptidases; Enzymes; Family; Future Generations; Genetic; Genetic Recombination; Genome; Goals; Grant; Health; Life; Malignant Neoplasms; Mechanics; Mediating; Methods; Modeling; N-acetylglucosamine-1-phosphodiester alpha-N-acetylglucosaminidase; N-terminal; Peptide Hydrolases; Peptide Signal Sequences; Peptides; Phase; Positioning Attribute; Process; Proteins; Proteolysis; Proteome; Proteomics; Reaction; Recycling; Role; Source; Specificity; Stress; Syndrome; Testing; Transposase; Work; design; environmental change; experience; interest; oxidation; protein structure; repaired; research study; response; unfoldase

DESCRIPTION (provided by applicant): Higher-order protein-DNA complexes both orchestrate and catalyze many of life's most central processes. Genetic recombination and DMA protection, 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 by AAA+ enzymes is a critical yet poorly understood aspect of the strategies used by cells to both interpret and protect their genomes.

描述（由申请人提供）：高阶蛋白质-DNA 复合物协调并催化许多生命最核心的过程。基因重组和 DMA 保护是维持子孙后代基因组完整性所必需的过程，尤其大量使用蛋白质-DNA 超结构。这些复合物的一个关键特征是它们通常非常稳定，因此可能需要 ATP 依赖性蛋白质解折叠酶和蛋白酶来重塑、拆除、破坏或回收组成蛋白质。该项目的重点是了解 Clp/Hsp100 ATPase 家族的蛋白质如何识别蛋白质复合物进行重塑或破坏。 Clp/Hsp100 蛋白是 AAA+ 酶的一个亚家族，利用 ATP 水解作用对其底物进行机械功。第一个具体目标是了解蛋白质解折叠酶 CIpX 如何识别促进噬菌体 Mu DNA 转座的蛋白质-DNA 复合物。提出了阐明转座酶内负责识别的肽信号的实验。此外，我们将测试一种模型，在该模型中，复合物的不对称特征引导 CIpX 的展开活性至一个特定的转座酶亚基，从而生成具有独特结构的新的（不太稳定）复合物。第二个目标是阐明 CIpX 如何识别 DNA 保护蛋白 Dps，以及如何调整这种识别以适应不断变化的环境条件。将研究肽信号、接头蛋白和 DNA 在 Dps 识别中的作用。最后，提出了蛋白质组学实验，旨在全面了解 ATP 依赖性蛋白质解折叠酶/蛋白酶的作用。这些实验将为特定衔接蛋白和肽结合域在 AAA+ 酶的底物选择中的作用提供蛋白质组范围的视图。保护基因组免受损伤并促进受损 DNA 忠实修复的细胞机制对于健康和生存至关重要，与这些细胞过程的遗传破坏相关的众多癌症综合征充分证明了这一点。 AAA+ 酶对蛋白质的重塑和破坏是细胞解释和保护其基因组所使用的策略的一个关键但人们知之甚少的方面。