Humana Cell Cycle Checkpoint Control Protocols: 241 (Methods in Molecular Biology, 241)

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. Review "...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 From the Back Cover 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.