Checkpoint Controls and Cancer: Checkpoint Controls and Cancer Activation and Regulation Protocols Volume 2

Checkpoint Controls and Cancer: Checkpoint Controls and Cancer Activation and Regulation Protocols Volume 2 : Volume 2: Activation and Regulation Protocols

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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.
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Product details

  • Hardback | 358 pages
  • 152.4 x 233.7 x 22.9mm | 748.44g
  • Totowa, NJ, United States
  • English
  • 2004 ed.
  • XVIII, 358 p.
  • 1588295001
  • 9781588295002

Back cover copy

Intracellular checkpoint controls constitute a network of signal transduction pathways that protect cells from external stresses and internal errors by means of cell cycle arrest, DNA repair, or apoptosis. Failure of this machinery can lead to embryonic death, genetic diseases, and cancer. In Checkpoint Controls and Cancer, Volume 2: Activation and Regulation Protocols, Axel H. Schönthal, PhD, marshals a prestigious panel of researchers working at the cutting edges of their fields to present readily reproducible experimental protocols for the study of the molecular components of checkpoint controls and their regulation. Described in step-by-step detail, these powerful techniques offer such novel approaches as the use of genome databases and siRNA to analyze how cells of the human body can escape proper surveillance to grow into a tumor. Additional experimental methods are provided for the manipulation of checkpoint pathways and the analysis of the resulting consequences for the cellular phenotype. The earlier first volume, Reviews and Model Systems, comprehensively describes the complexities of checkpoint controls and the model systems available to study them.
Comprehensive and up-to-date, the two volumes of Checkpoint Controls and Cancer offer novice and experienced researchers alike not only entré into the complexities of this vast field, but also to the full panoply of productive tools needed to deepen understanding of the systems, as well as to develop new and more effective therapies.
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Table of 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

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