Structural and Biochemical Analyses of Type II DNA Topoisomerases

II 型 DNA 拓扑异构酶的结构和生化分析

基本信息

批准号：
10112833
负责人：
JAMES M BERGER
金额：
$ 45.47万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-05-01 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10112833
关键词：
3-Dimensional ATP phosphohydrolase Address Adopted Anti-Bacterial Agents Antineoplastic Agents Architecture Area Binding Biochemical Biochemistry Biological Biology Catalytic Domain Cell Survival Cell physiology Cells Chemicals Chromosome Structures Chromosomes Clinical Color Communicable Diseases Complex DNA DNA Damage DNA topoisomerase II alpha Data Development Disease Drug Targeting Electron Microscopy Elements Enzymes Etiology Eukaryotic Cell Event Funding Gene Expression Genes Genetic Genetic Transcription Goals Health Homologous Gene Housekeeping Human Hybrids Image Knowledge Label Lead Left Ligand Binding Link Malignant Neoplasms Mass Spectrum Analysis Meiosis Meiotic Recombination Methodology Modeling Molecular Machines Mutation Outcome Pathway interactions Physiological Positioning Attribute Process Production Proteins Regulation Reporting Reproduction Research Route SPO11 gene Shapes Site Structure Superhelical DNA System Testing Therapeutic Intervention Topoisomerase Topoisomerase II Topoisomerase Inhibitors Transcriptional Activation Variant Work Yeasts base cancer cell dimer enzyme activity field study genetic information genome integrity in vivo inhibitor/antagonist insight molecular dynamics mutant paralogous gene programs promoter recombinase response scaffold

项目摘要

ABSTRACT The appropriate control of DNA topology has a major impact on the stability and flow of genetic information. The present application focuses on type II topoisomerases, molecular machines that modulate DNA supercoiling and remove chromosome entanglements by catalyzing the ATP-dependent transport of one DNA duplex through another. Type II topoisomerases are critical for maintaining gene expression, chromosome superstructure, and genome integrity; they also serve as frontline drug targets for treating infectious disease and cancer. During the prior project period, we gained several new insights into the mechanism, regulation, and DNA damage propensity of type II topoisomerases. These efforts open up three new research opportunities centered on complementary but non-interdependent aspects of type II topoisomerase action that will advance both fundamental biological knowledge and therapeutic intervention. Aim 1 seeks to explain the long-unresolved question of how type II topoisomerases link ATP-dependent allosteric responses to the selective engagement and release of multiple DNA segments, and to illuminate how a system responsible for initiating meiotic recombination co-opted and modified a topoisomerase scaffold to generate DNA breaks for promoting chromosome exchange. Aim 2 will break new ground in understanding the regulation of eukaryotic topo II, uncovering natural metabolites that control enzyme activity as means to modulate replication, transcription, and chromosome organization processes that are sensitive to DNA topology. Aim 3 will determine how the action of human topo IIβ drives aberrant DNA damaging events and how naturally-occurring mutations may further potentiate this detrimental activity. Our approach is distinguished by a comprehensive blend of biochemical, structural, computational, cell-based, and chemical biology methodologies. Past progress and unpublished findings provide data to establish feasibility for the proposed effort. Our studies will impact multiple fields, from the study of molecular machines and the control of DNA dynamics, to understanding how topoisomerase activity and its regulation support specific physiological needs and promote human reproduction and health.

抽象的 DNA拓扑结构的适当控制对DNA的稳定性和流动性有重大影响本申请集中于II型拓扑异构酶，调节 DNA 超螺旋并去除染色体的分子机器通过催化一个 DNA 双链体的 ATP 依赖性运输来实现缠结另一个是II型拓扑异构酶对于维持基因表达至关重要，染色体超结构和基因组完整性；它们也可作为一线药物治疗传染病和癌症的目标。在之前的项目期间，我们对该机制有了一些新的见解， II 型拓扑异构酶的调控和 DNA 损伤倾向。增加三个新的研究机会，重点是互补但不相互依赖 II 型拓扑异构酶作用的各个方面将促进基本生物学目标 1 试图解释长期悬而未决的问题。 II 型拓扑异构酶如何将 ATP 依赖性变构反应与多个 DNA 片段的选择性接合和释放，并阐明如何负责启动减数分裂重组的系统增选并修改了拓扑异构酶支架产生DNA断裂以促进染色体交换。目标 2 将在理解真核拓扑结构 II 的调控方面开辟新天地，发现控制酶活性的天然代谢物作为调节手段复制、转录和染色体组织过程对这些过程敏感 DNA 拓扑结构。目标 3 将确定人类拓扑 IIβ 的作用如何驱动异常。 DNA 损伤事件以及自然发生的突变如何进一步加剧这种情况活动。我们的方法以生化、结构、计算、基于细胞和化学生物学方法的过去的进展和。未发表的研究结果提供了数据来确定拟议工作的可行性。研究将影响多个领域，从分子机器的研究到控制 DNA 动力学，了解拓扑异构酶活性及其调节支持特定的生理需求并促进人类生殖和健康。

项目成果

期刊论文数量（47）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structure of the topoisomerase II ATPase region and its mechanism of inhibition by the chemotherapeutic agent ICRF-187.

拓扑异构酶 II ATP 酶区域的结构及其化疗药物 ICRF-187 的抑制机制。

DOI：
发表时间：
2003-09-16
期刊：
影响因子：
11.1
作者：
Classen, Scott;Olland, Stephane;Berger, James M
通讯作者：
Berger, James M

A domain insertion in Escherichia coli GyrB adopts a novel fold that plays a critical role in gyrase function.

大肠杆菌 GyrB 中的结构域插入采用了一种在旋转酶功能中发挥关键作用的新型折叠。

DOI：
发表时间：
2010-11
期刊：
影响因子：
14.9
作者：
Schoeffler, Allyn J;May, Andrew P;Berger, James M
通讯作者：
Berger, James M

Real-time detection of DNA topological changes with a fluorescently labeled cruciform.

利用荧光标记的十字形实时检测 DNA 拓扑变化。

DOI：
发表时间：
2013-07
期刊：
影响因子：
14.9
作者：
Jude, Kevin M;Hartland, Abbey;Berger, James M
通讯作者：
Berger, James M

Structural basis for the MukB-topoisomerase IV interaction and its functional implications in vivo.

MukB-拓扑异构酶 IV 相互作用的结构基础及其体内功能意义。

DOI：
发表时间：
2013-11-13
期刊：
影响因子：
0
作者：
Vos, Seychelle M;Stewart, Nichole K;Oakley, Martha G;Berger, James M
通讯作者：
Berger, James M

Design, synthesis and molecular docking of new N-4-piperazinyl ciprofloxacin-triazole hybrids with potential antimicrobial activity.

具有潜在抗菌活性的新型 N-4-哌嗪基环丙沙星-三唑杂化物的设计、合成和分子对接。

DOI：
10.1016/j.bioorg.2019.102952
发表时间：
2019-07-01
期刊：
Bioorganic chemistry
影响因子：
5.1
作者：
Hamada H. H. Mohammed;Elshimaa M N Abdelhafez;S. Abbas;Gamal A I Moustafa;G. Hauk;J. Berger;S. Mitarai;M. Arai;R. A. Abd El;G. Abuo‐Rahma
通讯作者：
G. Abuo‐Rahma