Cell Cycle Checkpoints: Methods and Protocols

Cell cycle checkpoints control the fidelity and orderly progression of eukaryotic cell division. By controlling the orderly progression of critical cell cycle events such as DNA replication and chromosome segregation and ensuring proper repair of damaged DNA, cell cycle checkpoints function to ensure genome integrity. Mechanisms of checkpoint controls are not only the research focus of investigators interested in mechanisms that regulate the cell cycle, but are also the interests of researchers studying cancer development as it is increasingly clear that loss of cell cycle checkpoints, which leads to genomic instability as a result, is a hallmark of tumorigenesis. Cell Cycle Checkpoints: Methods and Protocols provides detailed descriptions of methodologies currently employed by researchers in the field, including those commonly used in the mammalian, yeast, C. elegans, Drosophila, and Xenopus model systems. Each chapter describes a specific technique or protocol, such as a method to induce cell cycle checkpoints in a particular model system, to synchronize a population of cells to allow observations of cell cycle progression, to identify genes involved in checkpoint regulation, and to study particular protein components of cell cycle checkpoint pathways. Written in the highly successful Methods in Molecular Biology™ series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.   Authoritative and easily accessible, Cell Cycle Checkpoints: Methods and Protocols seeks to serve both professionals and novices with its well-honed methodologies in an effort to further our knowledge of this essentialfield.

Table of Contents:
Studying G2 DNA Damage Checkpoints using the Fission Yeast Schizosaccharomyces pombe.- Studying S-phase DNA Damage Checkpoints using the Fission Yeast Schizosaccharomyces pombe.- Methods for Studying the G2 DNA Damage Checkpoint in Mammalian Cells.- Evaluating Spindle Assembly Checkpoint Competence in Mouse Oocytes using Immunoblotting.- Analysis of Changes in Protein Level and Subcellular Localization during Cell Cycle Progression using the Budding Yeast Saccharomyces cerevisiae.- Studying Cell Cycle Checkpoints using Drosophila Cultured Cells.- In Vivo Live-Analysis of Cell Cycle Checkpoints in Drosophila Early Embryos.- Using Drosophila Larval Imaginal Discs to Study Low Dose Radiation-induced Cell Cycle Arrest.- Screening for Radiation Sensitizers of Drosophila Checkpoint Mutants.- Study of Cell Cycle Checkpoints using Xenopus Cell-free Extracts.- Analyzing DNA Replication Dynamics of Genotoxin-Treated Cells Using Velocity Sedimentation.- Methods For Studying Checkpoint Kinases – Chk1.- A Human Cell Extract-Based Assay for the Activation of ATM and ATR Checkpoint Kinases.- Analyzing p53 regulated DNA Damage Checkpoints by Flow Cytometry.- Using Drosophila S2 Cells to Measure S phase-Coupled Protein Destruction via Flow Cytometric Analysis.- Assessing G1-to-S-phase Progression After Genotoxic Stress.- Indirect Immunofluorescence for Monitoring Spindle Assembly and Disassembly in Yeast.- Detecting Recruitment of DNA Damage Response Factors through the eChIP Approach.- Methods to Study Cancer Therapeutic Drugs that Target Cell Cycle Checkpoints.