Cell cycle checkpoint control protocols
Author(s)
Bibliographic Information
Cell cycle checkpoint control protocols
(Methods in molecular biology / John M. Walker, series editor, v. 241)
Humana, c2004
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Note
Includes bibliographical references and index
Description and Table of Contents
Description
The field of cell cycle regulation is based on the observation that the life cycle of a cell progresses through several distinct phases, G1, M, S, and G2, occurring in a well-defined temporal order. Details of the mechanisms involved are rapidly emerging and appear extraordinarily complex. Furthermore, not only is the order of the phases important, but in normal eukaryotic cells one phase will not begin unless the prior phase is completed successfully. Che- point control mechanisms are essentially surveillance systems that monitor the events in each phase, and assure that the cell does not progress prematurely to the next phase. If conditions are such that the cell is not ready to progress-for example, because of incomplete DNA replication in S or DNA damage that may interfere with chromosome segregation in M-a transient delay in cell cycle progression will occur. Once the inducing event is properly handled- for example, DNA replication is no longer blocked or damaged DNA is repaired-cell cycle progression continues. Checkpoint controls have recently been the focus of intense study by investigators interested in mechanisms that regulate the cell cycle. Furthermore, the relationship between checkpoint c- trol and carcinogenesis has additionally enhanced interest in these cell cycle regulatory pathways. It is clear that cancer cells often lack these checkpoints and exhibit genomic instability as a result. Moreover, several tumor suppressor genes participate in checkpoint control, and alterations in these genes are as- ciated with genomic instability as well as the development of cancer.
Table of Contents
Part I. Induction and Detection of Changes in Cell Cycle Progression
Methods to Induce Cell Cycle Checkpoints
Howard B. Lieberman and Kevin M. Hopkins
Methods for Synchronizing Mammalian Cells
Michael H. Fox
Enrichment of Cells in Different Phases of the Cell Cycle by Centrifugal Elutriation
Tej K. Pandita
Analysis of the Mammalian Cell Cycle by Flow Cytometry
Haiying Hang and Michael H. Fox
Methods for Detecting Cells in S Phase
Wei-Hsin Sun and Melvin L. DePamphilis
Yeast Cell Synchronization
Audra Day, Colette Schneider, and Brandt L. Schneider
Analysis of the Budding Yeast Saccharomyces cerevisiae Cell Cycle by Morphological Criteria and Flow Cytometry
Hong Zhang and Wolfram Siede
Analysis of the Fission Yeast Schizosaccharomyces pombe Cell Cycle
Eliana B. Gomez and Susan L. Forsburg
Part II. Analysis of Genes Involved in Checkpoint Control
Strategies to Isolate Evolutionarily Conserved Cell Cycle Regulatory Genes
Scott Davey
Microarray Approaches for Analysis of Cell Cycle Regulatory Genes
Sally A. Amundson and Albert J. Fornace, Jr.
Using the Yeast Genome-Wide Gene-Deletion Collection for Systematic Genetic Screens
Jian Zhang, Lisa Ottmers, and Brandt L. Schneider
Gene Targeting in Cultured Human Cells
Todd A. Waldman
Use of In Vivo Gap Repair for Isolation of Mutant Alleles of a Checkpoint Gene
Migdalisel Colon and Nancy C. Walworth
In Vitro Mutagenesis to Define Functional Domains
Jian Qin, Zhe Peng, and Maureen V. McLeod
Use of Gene Overexpression to Assess Function in Cell Cycle Control
Erik K. Flemington and Antonio Rodriguez
Histone Acetylation/Deacetylation As a Regulator of Cell Cycle Gene Expression
Chenguang Wang, Maofu Fu, and Richard G. Pestell
Part III. Analysis ofProteins Involved in Checkpoint Control
Cataloging Proteins in Cell Cycle Control
Kazimierz O. Wrzeszczynski and Burkhard Rost
Multidimensional Proteomic Analysis of Proteolytic Pathways Involved in Cell Cycle Control
Michael W. Schmidt, Aruna Jain, and Dieter A. Wolf
Purification and Identification of Protein Complexes That Control the Cell Cycle
Matthew A. Burtelow, Vladimir N. Podust, and Larry M. Karnitz
Xenopus Cell-Free Extracts to Study DNA Damage Checkpoints
Vincenzo Costanzo and Jean Gautier
Protein-Protein Interactions
Graziella Pedrazzi and Igor Stagljar
Detection of Kinase and Phosphatase Activities
Sean M. Post and Eva Y.-H.P. Lee
Monitoring Changes in the Subcellular Location of Proteins in S. cerevisiae
Diego Rua, Teresa Holzen, Benjamin S. Glick, Stephen J. Kron, and Douglas K. Bishop
Part IV. Chromosomes and the Cell Cycle
Chromosomal Changes and Cell Cycle Checkpoints in Mammalian Cells
Charles R. Geard and Brian Ponnaiya
Detecting the Influence of Cell Cycle Regulatory Proteins on Human Telomeres
Tej K. Pandita
Monitoring Spindle Assembly and Disassembly in Yeast by Indirect Immunofluorescence
Rita K. Miller
Index
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