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Main description:
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.
Contents:
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
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 378
Weight: 575g
Availability: Available
Subcategories: Oncology
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