Cell Cycle Control and Dysregulation Protocols

by Giordano, Antonio; Romano, Gaetano

• ISBN13: 9780896039490
• ISBN10: 0896039498
• Format: Hardcover
• Copyright: 2004-08-01
• Publisher: Humana Pr Inc

Summary

Expert laboratorians detail emerging methodologies for studying the cell cycle, its kinases, and kinase inhibitors. The authors focus the issues of gene expression in vivo and in vitro, the analysis of cyclin-dependent kinase inhibitors, protein degradation mediated by the proteosome, the analysis of the transformed cell phenotype, and innovative techniques to detect apoptosis. Many of the protocols are new, such as electron microsocopy to detect apoptosis and proteosome-mediated degradation, whereas others study the interactions between protein-protein, protein-DNA, and protein-RNA.

Table of Contents

Preface	v
Contributors	xi
Part I. Analysis of Cyclins and Cyclin-Dependent Kinases
The Biology of Cyclins and Cyclin-Dependent Protein Kinases: An Introduction	3	(20)
Lucio Miele
In Situ Immunofluorescence Analysis: Immunofluorescence Microscopy	23	(6)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
In Situ Immunofluorescence Analysis: Analyzing RNA Synthesis by 5-Bromouridine-5'-Triphosphate Labeling	29	(4)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Immunofluorescence Analysis Using Epitope-Tagged Proteins: In Vitro System	33	(4)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Analysis of In Vivo Gene Expression Using Epitope-Tagged Proteins	37	(4)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Chromatin Immunoprecipitation	41	(4)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression: Electrophoretic Mobility Shift Assay	45	(12)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression: DNase I Digestion	57	(6)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Protein-Deoxyribonucleic Acid Interactions Linked to Gene Expression: Ligation-Mediated Polymerase Chain Reaction	63	(6)
Amjad Javed
Sayyed K. Zaidi
Soraya E. Gutierrez
Christopher J. Lengner
Kimberly S. Harrington
Hayk Hovhannisyan
Brian C. Cho
Jitesh Pratap
Shirwin M. Pockwinse
Martin Montecino
Andre J. van Wijnen
Jane B. Lian
Janet L. Stein
Gary S. Stein
Assays for Cyclin-Dependent Kinase Inhibitors	69	(10)
Adrian M. Senderowicz
Protein Degradation Via the Proteosome	79	(16)
Henry Hoff
Hong Zhang
Christian Sell
Part II. Analysis of the Factors Involved in Cell Cycle Deregulation
The Transformed Phenotype	95	(10)
Henry Hoff
Barbara Belletti
Hong Zhang
Christian Sell
A Morphologic Approach to Detect Apoptosis Based on Electron Microscopy	105	(8)
Martyn K. White
Caterina Cinti
Detection of Apoptotic Deoxyribonucleic Acid Break by In Situ Nick Translation	113	(8)
Carmela Trimarchi
Dario La Sala
Alessandra Zamparelli
Caterina Cinti
Part III. Cellular Response to Deoxyribonucleic Acid Damage
Induction of Deoxyribonucleic Acid Damage by Alkylating Agents	121	(6)
Salvatore Cortellino
David P. Turner
Domenico Albino
Alfonso Bellacosa
Induction of Deoxyribonucleic Acid Damage by γ Irradiation	127	(6)
Salvatore Cortellino
David P. Turner
Alfonso Bellacosa
Ultraviolet Irradiation of Cells	133	(8)
David P. Turner
Anthony T. Yeung
Alfonso Bellacosa
Part IV. Retroviridae-Based Vectors: Protocols for Lentiviral- and Retroviral-Mediated Gene Transfer to Engineer Cell Culture Systems
Transient Production of Retroviral- and Lentiviral-Based Vectors for the Transduction of Mammalian Cells	141	(8)
Tiziana Tonini
Pier Paolo Claudio
Antonio Giordano
Gaetano Romano
Determination of Functional Viral Titer by Drug-Resistance Colony Assay, Expression of Green Fluorescent Protein, and β-Galactoside Staining	149	(6)
Tiziana Tonini
Pier Paolo Claudio
Antonio Giordano
Gaetano Romano
Retroviral and Lentiviral Vector Titration by the Analysis of the Activity of Viral Reverse Transcriptase	155	(6)
Tiziana Tonini
Pier Paolo Claudio
Antonio Giordano
Gaetano Romano
Part V. Detection of Gene Expression in Subcellular Compartments
Single and Double Colloidal Gold Labeling in Postembedding Immunoelectron Microscopy	161	(10)
Nicoletta Zini
Liliana Solimando
Caterina Cinti
Nadir Mario Maraldi
Multifluorescence Labeling and Colocalization Analyses	171	(8)
Massimo Riccio
Maja Dembic
Caterina Cinti
Spartaco Santi
Index	179