Mechanisms of DNA Recombination and Genome Rearrangements: Methods to Study Homologous Recombination, Volume 600, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field.
Homologous genetic recombination remains the most enigmatic process in DNA metabolism. The molecular machines of recombination preserve the integrity of the genetic material in all organisms and generate genetic diversity in evolution. The same molecular machines that support genetic integrity by orchestrating accurate repair of the most deleterious DNA lesions, however, also promote survival of cancerous cells and emergence of radiation and chemotherapy resistance. This two-volume set offers a comprehensive set of cutting edge methods to study various aspects of homologous recombination and cellular processes that utilize the enzymatic machinery of recombination The chapters are written by the leading researches and cover a broad range of topics from the basic molecular mechanisms of recombinational proteins and enzymes to emerging cellular techniques and drug discovery efforts.
1. Cover image - p. iv 2. Title page - p. iv 3. Table of Contents - p. iv 4. Copyright - p. iv 5. Contributors - p. xiii 6. Preface - p. xvii 7. Chapter One: Processing of DNA Double-Strand Breaks in Yeast - p. xviii 8. Chapter Two: Methods to Study DNA End Resection I: Recombinant Protein Purification - p. 24 9. Chapter Three: Methods to Study DNA End Resection II: Biochemical Reconstitution Assays - p. 66 10. Chapter Four: Direct Quantitative Monitoring of Homology-Directed DNA Repair of Damaged Telomeres - p. 106 11. Chapter Five: Kinetic Analysis of the Exonuclease Activity of the Bacteriophage T4 Mre11–Rad50 Complex - p. 134 12. Chapter Six: Expression, Purification, and Biochemical Evaluation of Human RAD51 Protein - p. 156 13. Chapter Seven: Determining the RAD51-DNA Nucleoprotein Filament Structure and Function by Cryo-Electron Microscopy - p. 178 14. Chapter Eight: Observation and Analysis of RAD51 Nucleation Dynamics at Single-Monomer Resolution - p. 199 15. Chapter Nine: TIRF-Based Single-Molecule Detection of the RecA Presynaptic Filament Dynamics - p. 232 16. Chapter Ten: The RadA Recombinase and Paralogs of the Hyperthermophilic Archaeon Sulfolobus solfataricus - p. 253 17. Chapter Eleven: Reconstituting the 4-Strand DNA Strand Exchange - p. 284 18. Chapter Twelve: Purification of Saccharomyces cerevisiae Homologous Recombination Proteins Dmc1 and Rdh54/Tid1 and a Fluorescent D-Loop Assay - p. 305 19. Chapter Thirteen: Probing Dynamic Assembly and Disassembly of Rad51 Tuned by Srs2 Using smFRET - p. 320 20. Chapter Fourteen: The Recombination Mediator BRCA2: Architectural Plasticity of Recombination Intermediates Revealed by Single-Molecule Imaging (SFM/TIRF) - p. 345 21. Chapter Fifteen: Single-Molecule Dynamics and Localization of DNA Repair Proteins in Cells - p. 374 22. Chapter Sixteen: Single-Stranded DNA Curtains for Studying the Srs2 Helicase Using Total Internal Reflection Fluorescence Microscopy - p. 406 23. Chapter Seventeen: Single-Molecule Analysis of Replication Protein A–DNA Interactions - p. 437 24. Chapter Eighteen: Single-Molecule Studies of ssDNA-Binding Proteins Exchange - p. 461 25. Chapter Nineteen: Dissecting the Recombination Mediator Activity of BRCA2 Using Biochemical Methods - p. 477 26. Chapter Twenty: Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis - p. 511 27. Chapter Twenty-One: GEN1 Endonuclease: Purification and Nuclease Assays - p. 525 28. Chapter Twenty-Two: Biochemical and Structural Properties of Fungal Holliday Junction-Resolving Enzymes - p. 542 29. Chapter Twenty-Three: Preparation and Resolution of Holliday Junction DNA Recombination Intermediates - p. 568
Mechanisms of DNA Recombination and Genome Rearrangements: Methods to Study Homologous Recombination, Volume 600, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field.
Homologous genetic recombination remains the most enigmatic process in DNA metabolism. The molecular machines of recombination preserve the integrity of the genetic material in all organisms and generate genetic diversity in evolution. The same molecular machines that support genetic integrity by orchestrating accurate repair of the most deleterious DNA lesions, however, also promote survival of cancerous cells and emergence of radiation and chemotherapy resistance. This two-volume set offers a comprehensive set of cutting edge methods to study various aspects of homologous recombination and cellular processes that utilize the enzymatic machinery of recombination The chapters are written by the leading researches and cover a broad range of topics from the basic molecular mechanisms of recombinational proteins and enzymes to emerging cellular techniques and drug discovery efforts.
1. Cover image - p. iv 2. Title page - p. iv 3. Table of Contents - p. iv 4. Copyright - p. iv 5. Contributors - p. xiii 6. Preface - p. xvii 7. Chapter One: Processing of DNA Double-Strand Breaks in Yeast - p. xviii 8. Chapter Two: Methods to Study DNA End Resection I: Recombinant Protein Purification - p. 24 9. Chapter Three: Methods to Study DNA End Resection II: Biochemical Reconstitution Assays - p. 66 10. Chapter Four: Direct Quantitative Monitoring of Homology-Directed DNA Repair of Damaged Telomeres - p. 106 11. Chapter Five: Kinetic Analysis of the Exonuclease Activity of the Bacteriophage T4 Mre11–Rad50 Complex - p. 134 12. Chapter Six: Expression, Purification, and Biochemical Evaluation of Human RAD51 Protein - p. 156 13. Chapter Seven: Determining the RAD51-DNA Nucleoprotein Filament Structure and Function by Cryo-Electron Microscopy - p. 178 14. Chapter Eight: Observation and Analysis of RAD51 Nucleation Dynamics at Single-Monomer Resolution - p. 199 15. Chapter Nine: TIRF-Based Single-Molecule Detection of the RecA Presynaptic Filament Dynamics - p. 232 16. Chapter Ten: The RadA Recombinase and Paralogs of the Hyperthermophilic Archaeon Sulfolobus solfataricus - p. 253 17. Chapter Eleven: Reconstituting the 4-Strand DNA Strand Exchange - p. 284 18. Chapter Twelve: Purification of Saccharomyces cerevisiae Homologous Recombination Proteins Dmc1 and Rdh54/Tid1 and a Fluorescent D-Loop Assay - p. 305 19. Chapter Thirteen: Probing Dynamic Assembly and Disassembly of Rad51 Tuned by Srs2 Using smFRET - p. 320 20. Chapter Fourteen: The Recombination Mediator BRCA2: Architectural Plasticity of Recombination Intermediates Revealed by Single-Molecule Imaging (SFM/TIRF) - p. 345 21. Chapter Fifteen: Single-Molecule Dynamics and Localization of DNA Repair Proteins in Cells - p. 374 22. Chapter Sixteen: Single-Stranded DNA Curtains for Studying the Srs2 Helicase Using Total Internal Reflection Fluorescence Microscopy - p. 406 23. Chapter Seventeen: Single-Molecule Analysis of Replication Protein A–DNA Interactions - p. 437 24. Chapter Eighteen: Single-Molecule Studies of ssDNA-Binding Proteins Exchange - p. 461 25. Chapter Nineteen: Dissecting the Recombination Mediator Activity of BRCA2 Using Biochemical Methods - p. 477 26. Chapter Twenty: Approaches to Understanding the Mediator Function of Brh2 in Ustilago maydis - p. 511 27. Chapter Twenty-One: GEN1 Endonuclease: Purification and Nuclease Assays - p. 525 28. Chapter Twenty-Two: Biochemical and Structural Properties of Fungal Holliday Junction-Resolving Enzymes - p. 542 29. Chapter Twenty-Three: Preparation and Resolution of Holliday Junction DNA Recombination Intermediates - p. 568
