CiNii 図書 - Activation and regulation protocols

Intracellular checkpoint controls constitute a network of signal transd- tion pathways that protect cells from external stresses and internal errors. Ext- nal stresses can be generated by the continuous assault of DNA-damaging agents, such as environmental mutagens, ultraviolet (UV) light, ionizing radiation, or the reactive oxygen species that can arise during normal cellular metabolism. In response to any of these assaults on the integrity of the genome, the activation of the network of checkpoint control pathways can lead to diverse cellular responses, such as cell cycle arrest, DNA repair, or elimination of the cell by cell death (apoptosis) if the damage cannot be repaired. Moreover, internal errors can occur during the highly orchestrated replication of the cellular genome and its distribution into daughter cells. Here, the temporal order of these cell cycle events must be strictly enforced-for example, to ensure that DNA replication is c- plete and occurs only once before cell division, or to monitor mitotic spindle assembly, and to prevent exit from mitosis until chromosome segregation has been completed. Thus, well functioning checkpoint mechanisms are central to the maintenance of genomic integrity and the basic viability of cells and, the- fore, are essential for proper development and survival. The importance of proper functioning of checkpoints becomes plainly obvious under conditions in which this control network malfunctions and fails. Depending on the severity and timing, failure of this machinery can lead to embryonic lethality, genetic diseases, and cancer.

Part I. Protocols for the Study of Checkpoint-Regulatory Components Analysis of RB Action in DNA Damage Checkpoint Response Christopher N. Mayhew, Emily E. Bosco, David A. Solomon, Erik S. Knudsen, and Steven P. Angus Interaction Between the Retinoblastoma Protein and Protein Phosphatase 1 During the Cell Cycle Norbert Berndt and John W. Ludlow Generation of p53 Target Database Via Integration of Microarray and Global p53 DNA-Binding Site Analysis Suxing Liu, Asra Mirza, and Luquan Wang Functional Analysis of CDK Inhibitor p21WAF1 Rati Fotedar, Mourad Bendjennat, and Arun Fotedar Analysis of p21CDKN1A Recruitment to DNA Excision Repair Foci in the UV-Induced DNA Damage Response Lucia A. Stivala and Ennio Prosperi Quantitative Determination of p16 Gene Expression by RT-PCR Sylke Schneider, Kazumi Uchida, Dennis Salonga, Ji Min Yochim, Kathleen D. Danenberg, and Peter V. Danenberg Measuring Cyclin-Dependent Kinase Activity Axel H. Schoenthal Determination of the Catalytic Activities of mTOR and Other Members of the Phosphoinositide-3-Kinase-Related Kinase Family Gary G. Chiang and Robert T. Abraham CHK1 Kinase Activity Assay Ya Wang and Hongyan Wang Assaying Cdc25 Phosphatase Activity Ingo Hassepass and Ingrid Hoffmann Analyzing the Regulation and Function of ATM Martin F. Lavin, Shaun P. Scott, Sergei Kozlov, and Nuri Gueven Use of siRNA to Study the Function of MDC1 in DNA Damage Responses Zhenkun Lou and Junjie Chen Functional Analysis of APC-Cdh1 Tamotsu Sudo, Naoto T. Ueno, and Hideyuki Saya Purification of Mitotic Checkpoint Complex, an Inhibitor of the APC/C From HeLa Cells Valery Sudakin and Tim J. Yen Analysis of the Spindle-Assembly Checkpoint in HeLa Cells Paul R. Andreassen, Dimitrios A. Skoufias, and Robert L.Margolis Functional Analysis of the Spindle-Checkpoint Proteins Using an In Vitro Ubiquitination Assay Zhanyun Tang and Hongtao Yu Part II. Studying Consequences of Checkpoint Pathway Activation Analysis of Checkpoint Responses to Histone Deacetylase Inhibitors Heather Beamish, Robyn Warrener, and Brian G. Gabrielli Biochemical Analysis of the Cell Cycle and Cell Cycle Checkpoints in Transiently Transfected Cells After Collection With Magnetic Beads Xiaofen Ye, Maxim Poustovoitov, Hidelita Santos, David M. Nelson, and Peter D. Adams Analysis of DNA Repair and Chromatin Assembly In Vitro Using Immobilized Damaged DNA Substrate Jill A. Mello, Jonathan G. Moggs, and Genevieve Almouzni Analyzing Cell Cycle Checkpoints After Ionizing Radiation Bo Xu and Michael B. Kastan FACS-Based Detection of Phosphorylated Histone H3 for the Quantitation of Mitotic Cells William R. Taylor Analysis of Cell Cycle by Flow Cytometry Piotr Pozarowski and Zbigniew Darzynkiewicz Analyzing Markers of Apoptosis In Vitro Stephanie Plenchette, Rodolphe Filomenko, Emmanuelle Logette, Stephanie Solier, Nelly Buron, Severine Cathelin, and Eric Solary Analysis of Telomerase Activity and Telomere Function in Cancer Katrina E. Gordon and E. Kenneth Parkinson Index

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Activation and regulation protocols

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