Mechanisms of DNA Recombination and Genome Rearrangements: Methods to Study Homologous Recombination - 1st Edition - ISBN: 9780128144299

Mechanisms of DNA Recombination and Genome Rearrangements: Methods to Study Homologous Recombination, Volume 600

1st Edition

Serial Volume Editors: Maria Spies Anna Malkova
Hardcover ISBN: 9780128144299
Imprint: Academic Press
Published Date: 1st March 2018
Page Count: 412
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Table of Contents

1. DSB processing in yeast
Loraine Symington
2. Purification of DNA end resection enzymes
Petr Ceijka
3. Biochemistry of DNA end resection
Petr Ceijka
4. Visualization of telomere movement and interchromosomal homology searches in ALT
Roger Greenberg
5. Pre-steady state kinetic methods for investigating the DNA exonuclease activity of Mre11
Scott Nelson
6. Production of large quantities of human RAD51
Maria Spies
7. Determining the RAD51 nucleoprotein filamentous structure and mechanism by CryoEM
Hong-Wei Wang
8. Single-molecule TIRF analysis of human RAD51 nucleation
Maria Spies
9. TIRF-based single-molecule detection of the RecA presynaptic filament dynamics
Sung Hyun Kim
10. Archaeal (Sulpholobus) RadA and its paralogs
Cynthia Haseltine
11. 4-strand DNA strand exchange
Alexander Mazin
12. Purification and biochemistry of DMC1 recombinase
Doug Bishop
13. Single-molecule TIRF analysis of yeast Rad51 nucleoprotein disassembly by helicases
Sua Myong
14. The recombination mediator BRCA2: architectural plasticity of recombination intermediates revealed by single molecule imaging (SFM/TIRF)
Claire Wyman
15. Visualisation and quantitation of BRCA2/RAD51 dynamics in cells
Roland Kanaar
16. RPA - DNA curtains
Eric Greene
17. Single-molecule studies of ssDNA binding proteins
Taekjip Ha
18. BRCA2 as recombination mediator
Aura Carreira
19. Ustilago maydis Brh2 as a model for human BRCA2
Bill Holloman
20. Purification and analysis of holiday junction resolvases
Stephen West
21. Structural analysis of HJ resolving enzyme GEN1
David Lilley
22. The preparation of four-stranded recombination intermediates for use in the HJ resolution reactions 
Stephen West


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.

Key Features

  • Contributions by the leading experts in the field of DNA repair, recombination, replication and genome stability
  • Documents cutting edge methods


Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists


No. of pages:
© Academic Press 2018
Academic Press
Hardcover ISBN:


Praise for the Series:
"Should be on the shelves of all libraries in the world as a whole collection." --Chemistry in Industry
"The work most often consulted in the lab." --Enzymologia
"The Methods in Enzymology series represents the gold-standard." --Neuroscience

Ratings and Reviews

About the Serial Volume Editors

Maria Spies Serial Volume Editor

Graduate of Peter the Great St. Petersburg Polytechnic University, Russia (1996 MS diploma with honors (equivalent of cum laude) in physics/biophysics) and Osaka University, Japan (2000 PhD in biological sciences), Dr. Maria Spies is an Associate Professor of Biochemistry at the University of Iowa Carver College of Medicine. Spies’ research career has been focused on deciphering the intricate choreography of the molecular machines orchestrating the central steps in the homology directed DNA repair. Her doctoral research supported by the Japanese Government (MONBUSHO) Graduate Scholarship provided the first detailed biochemical characterization of archaeal recombinase RadA. In her postdoctoral work with Dr. Steve Kowalczykowski (UC Davis) supported by the American Cancer Society, Spies reconstituted at the single-molecule level the initial steps of bacterial recombination and helped to explain how this process is regulated. Spies’ laboratory at the University of Iowa emphasizes the molecular machinery of homologous recombination, how it is integrated into DNA replication, repair and recombination (the 3Rs of genome stability), and how it is misappropriated in the molecular pathways that process stalled DNA replication events and DNA breaks through highly mutagenic, genome destabilizing mechanisms. Her goal is to understand, reconstitute and manipulate an elaborate network of DNA recombination, replication and repair, and to harness this understanding for anticancer drug discovery. The Spies lab utilizes a broad spectrum of techniques from biochemical reconstitutions of the key biochemical reactions in DNA recombination, repair and replication, to structural and single-molecule analyses of the proteins and enzymes coordinating these reactions, to combined HTS/CADD campaigns targeting human DNA repair proteins. Work in Spies Lab has been funded by the American Cancer Society (ACS), Howard Hughes Medical Institute (HHMI), and is currently supported by the National Institutes of Health (NIH). She received several prestigious awards including HHMI Early Career Scientist Award and Margaret Oakley Dayhoff Award in Biophysics. She serves on the editorial board of the Journal of Biological Chemistry, and as an academic editor of the journal Plos-ONE. She is a permanent member and a chair of the American Cancer Society “DNA mechanisms in cancer” review panel.

Affiliations and Expertise

Carver College of Medicine, University of Iowa, USA

Anna Malkova Serial Volume Editor

Graduate of the St. Petersburg State University, Russia (1987 MS diploma with honors (equivalent of cum laude) in Genetics and 1993 Ph.D. in Genetics), Dr. Anna Malkova is an Associate Professor of Biology Department at the University of Iowa College of Liberal Arts and Sciences. Malkova’s research is focused on DNA repair mechanisms. She investigates the repair of double-strand DNA breaks (DSBs), the most lethal type of DNA lesions. Using a dependable and powerful model system in yeast, where a single DSB is initiated by a site-specific HO endonuclease, she successfully demonstrated that imprecise or faulty repair of DSBs leads to structural genomic variations including mutations, copy number variations (CNVs), and chromosomal rearrangements similar to those that cause genetic diseases and cancer in humans. She has a longstanding interest in break-induced replication (BIR), a DSB repair pathway that repairs so-called one-ended DNA breaks, similar to those produced at eroded telomeres and by collapsed replication forks. In her postdoctoral work with Dr. James Haber (Brandeis University), she was among the first who identified this mechanism in eukaryotes. Studies in Malkova’s laboratory provided important insights into the mechanism of BIR, which was found to be fundamentally different from the mechanism of S-phase DNA replication. In particular, it was discovered that BIR is carried out by a migrating bubble rather than by a replication fork, and leads to conservative inheritance of newly synthesized DNA. This unusual molecular mechanism explained bursts of genetic instability that were observed in association with BIR, and that were similar to those associated with cancer and other diseases rooted in genetic instability. Work in Malkova’s lab is supported by the National Institutes of Health (NIH). Anna serves as a member to the Cancer Etiology study section of NIH/NCI.

Affiliations and Expertise

Department of Biology, University of Iowa, Iowa City, Iowa, USA