Cell Cycle Checkpoint Control Protocols

ISBN-13: 9781588291158 / Angielski / Twarda / 2003 / 376 str.

Howard B. Lieberman; Howard B. Lieberman

Opis

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."

Recenzje

"...includes step-by-step instructions written by an investigator who performs it frequently, an introduction explaining the principle behind the method, equipment and reagent lists, and tips on troubleshooting and avoiding known pitfalls." - Tumori

Spis treści

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. Gómez 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 Colòn 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

Blurb

In recent years, cell cycle checkpoints, cellular mechanisms that control the cell cycle and ensure genomic stability, have emerged as significant factors in carcinogenesis and in cancer cells. In Cell Cycle Checkpoint Control Protocols, leading investigators present their best methodologies to probe the mechanisms underlying cell cycle regulation and checkpoint control. Using mammalian, yeast, and frog systems, these experts describe readily reproducible methods to induce cell cycle checkpoints, detect changes in cell cycle progression, identify and analyze genes and proteins that regulate the process, and characterize chromosomal status as a function of cell cycle phase and progression. Each fully tested technique includes step-by-step instructions written by an investigator who performs it frequently, an introduction explaining the principle behind the method, equipment and reagent lists, and tips on troubleshooting and avoiding known pitfalls. Taken as a whole, the collection describes the major methodologies used by researchers in the field.
Cutting-edge and highly practical, Cell Cycle Checkpoint Control Protocols provides an extensive array of detailed protocols by which both experienced and novice investigators may successfully illuminate questions concerning cell cycle control.