DNA replication in eukaryotes is an important field, particular ly because of its direct impact on the study of cancer. The under standing of molecular mechanisms of replication and their regulation should allow a better comprehension of the alterations that lead to the proliferation of tumor cells and to error-prone repair in cells exposed to radiation or chemical carcinogens. During the last several years, many enzymes and proteins which participate in replication of DNA in eukaryotic cells have been identified, isolated and characterized. New concepts in chromatin structure have refocused attention on the study of replication of DNA complexed with his tones and non-histone chromosomal proteins. However, progress has been noticeably slower than for prokaryotes, essentially because of the difficulty in genetic analysis of eukaryo tic DNA replication. In June 1980, a workshop was organized in Cargese, Corsica (France) to facilitate exchanges of information between workers spe cializing in prokaryotes and those specializing in eukaryotes, and to allow discussion of new experimental approaches. With this in mind, special interest has been taken in the origin and termination of chromosome cycles and how they are controlled.

This book reviews the latest trends and future directions of DNA replication research. The contents reflect upon the principles that have been established through the genetic and enzymatic studies of bacterial, viral, and cellular replication during the past decades. The book begins with a historical overview of the studies on eukaryotic DNA replication by Professor Thomas Kelly, a pioneer of the field. The following chapters include genome-wide studies of replication origins and initiation factor binding, as well as the timing of DNA replications, mechanisms of initiation, DNA chain elongation and termination of DNA replication, the structural basis of functions of protein complexes responsible for execution of DNA replication, cell cycle-dependent regulation of DNA replication, the nature of replication stress and cells’ strategy to deal with the stress, and finally how all these phenomena are interconnected to genome instability and development of various diseases. By reviewing the existing concepts ranging from the old principles to the newest ideas, the book gives readers an opportunity to learn how the classical replication principles are now being modified and new concepts are being generated to explain how genome DNA replication is achieved with such high adaptability and plasticity. With the development of new methods including cryoelectron microscopy analyses of huge protein complexes, single molecular analyses of initiation and elongation of DNA replication, and total reconstitution of eukaryotic DNA replication with purified factors, the field is enjoying one of its most exciting moments, and this highly timely book conveys that excitement to all interested readers.

Eukaryotic DNA Replication: A Practical Approach is a comprehensive practical manual, with each of its eleven chapters describing an aspect of the methods currently used to investigate DNA replication in eukaryotes. The sequence of the chapters corresponds roughly to the order of events during DNA replication. The first chapters are concerned with initiation, looking at methods to characterize origins of replication and the proteins that interact with them. There then follow chapters describing protocols for the study of the elongation phase and the synthesis of the telomeres. The final chapters provide a more general overview of the study of DNA replication - including its investigation in model systems such as yeast, xenopus and viruses, and looks into methods used to study DNA:protein interactions that could be applied to the study of replication proteins. This exciting new volume provides over 120 tried and tested protocols for the analysis of eukaryotic DNA replication and will be of major interest to a wide variety of molecular and cell biologists, biochemists and medical researchers.

Replication-Coupled Repair, Volume 661 in the Methods in Enzymology series, highlights new advances in the field, with this new volume presenting interesting chapters on a variety of timely topics, including the Repair of replication-born DNA breaks by sister chromatid recombination, High resolution and high throughput DNA cyclization measurements to interrogate DNA bendability, A programmable detection method for genomic signatures: from disease diagnosis to genome editing, Characterization of the telomerase modulating activities of yeast DNA helicases, Eukaryotic DNA replication with purified budding yeast proteins, Single molecule studies of yeast Rad51 paralogs, Light activation and deactivation of Cas9 for DNA repair studies, and more. Other chapters explore MIDAS: Direct sequencing to map mitotic DNA synthesis and common fragile sites at high precision, Studying the DNA damage response in embryonic systems, GLASS-ChIP to map Mre11 cleavage sites in the human genome, New chemical biology approaches to trap reaction intermediates in living cells, Single-molecule imaging approaches for monitoring replication fork conflicts at genomic DNA G4 structures and R-loops in human cells, Monitoring the replication of structured DNA through heritable epigenetic change, Visualizing replication fork encounters with DNA interstrand crosslinks, and much more. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in Methods in Enzymology series Includes the latest information on replication-coupled repair

Written by leading experts, this learned but accessible book highlights the latest work on eukaryotic DNA replication.