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Main description:
Beginning with the Escherichia coli ? protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes have been identified that catalyze changes in the linkage of DNA strands. DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most p- cesses involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents. Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytotoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents. The two volumes of DNA Topoisomerase Protocols are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental protocols. The topics covered in Part I: DNA Topology and Enzymes, range from detailed methods to analyze various aspects of DNA structure, from linking number, knotting/unknotting, site-specific recombi- tion, and decatenation to the overexpression and purification of bacterial and eukaryotic DNA topoisomerases from a variety of cell systems and tissues.
Contents:
Assaying DNA Topoisomerase I Relaxation Activity
Lance Stewart and James J. Champoux
DNA Topoisomerase II-Catalyzed DNA Decatenation
Andrea Haldane and Daniel M. Sullivan
Plasmid DNA Supercoiling by DNA Gyrase
Penny J. Sayer, Martin L. Goble, Mark Oram, and L. Mark Fisher
Analyzing Reverse Gyrase Activity
Michel Duguet, Christine Jaxel, Anne-Ceile Declais, Fabrice Confalonieri, Janine Marsault, Claire Bouthier de la Tour, Marc Nadal, and Christiane Portemer
Topoisomerase II-Catalyzed ATP Hydrolosis as Monitored by Thin-Layer Chromatography
Paul S. Kingma, John M. Fortune, and Neil Osheroff
Use of a Real-Time, Coupled Assay to Measure the ATPase Activity of DNA Topoisomerase II
Janet E. Lindsley
Analysis of Topoisomerase-DNA Interactions by Electrophoretic Mobility Shift Assay
Stewart Shuman
Filter Binding Assays for Topoisomerase-DNA Complexes
Joaquim Roca
DNA Topoisomerase I-Mediated Nicking of Circular Duplex DNA
James J. Champoux
Analysis of Cleavage Complexes Using Reactive Inhibitor Derivatives
Kenneth N. Kreuzer, Catherine H. Freudenreich, and Yves Pommier
Uncoupling of Topoisomerase-Mediated DNA Cleavage and Religation
Anni H. Anderson, Kent Christiansen, and Ole Westergaard
Synthesis and Use of DNA Containing a 5'-Bridging Phosphorothioate as a Suicide Substrate for Type I DNA Topoisomerases
Alex B. Burgin, Jr
Isolation of Covalent Enzyme-DNA Complexes
Thomas C. Rowe, Dale Grabowski, and Ram Ganapathi
ICE Bioassay: Isolating In Vivo Complexes of Enzyme to DNA
Deepa Subramanian, Christine Sommer Furbee, and Mark T. Muller
DNA Unwinding Test Using Eukaryotic DNA Topoisomerase I
Linus L. Shen
Drug-DNA Interactions
David E. Graves
Quinolone Interactions with DNA and DNA Gyrase
Linus L.Shen
Bactericidal Assays for Fluoroquinolones
Thomas D. Gootz
Drug Toxicity in E. coli Cells Expressing Human DNA Topoisomerase I
Scott T. Taylor and Rolf Menzel
Drug-Induced Cytotoxicity in Tissue Culture
Cynthia E. Herzog and Leonard A. Zwelling
Analysis of Drug Uptake-Flow Cytometry
Grace Zoorob and Thomas Burke
Cell Cycle Analysis of Drug-Treated Cells
Frank Traganos, Gloria Juan, and Zbigniew Darzynkiewicz
Assaying Drug-Induced Apoptosis
Zbigniew Darzynkiewicz, Gloria Juan, and Frank Traganos
Single-Strand Conformational Polymorphism Analysis of DNA Topoisomerases I and II
Mary K. Danks
PCR-Based Cloning of DNA Topoisomerase Genes
Wai Mun Huang
Topoisomerase II-Catalyzed Relaxation and Catenation of Plasmid DNA
John M. Fortune and Neil Osheroff
Topoisomerase II-Mediated Cleavage of Plasmid DNA
D. Andrew Burden, Stacie J. Froelich-Ammon, and Neil Osheroff
Drug-induced Stabilization of Covalent DNA Topoisomerase I-DNA Intermediates: DNA Cleavage Assays
Paola Fiorani, Christine L. Hann, Piero Benedetti, and Mary-Ann Bjornsti
Studying DNA Topoisomerase I-Targeted Drugs in the Yeast Saccharomyces cerevisiae
Michael H. Woo, John R. Vance, and Mary-Ann Bjornsti
Yeast Systems for Demonstrating the Transfer of Antitopoisomerase II Agents
John L. Nitiss and Karen C. Nitiss
Index
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 348
Weight: 512g
Availability: Available
Subcategories: Pharmacology
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