ATM Kinase

1. Front Matter

   Pages i-xiv

   PDF

2. Assaying Radiosensitivity of Ataxia-Telangiectasia

   • Hailiang Hu, Shareef Nahas, Richard A. Gatti

   Pages 1-11

3. Assaying for Radioresistant DNA Synthesis, the Hallmark Feature of the Intra-S-Phase Checkpoint Using a DNA Fiber Technique

   • Amanda W. Kijas, Martin F. Lavin

   Pages 13-23

4. ATM Gene Mutation Detection Techniques and Functional Analysis

   • Guillaume Rieunier, Catherine Dubois D’Enghien, Alice Fievet, Dorine Bellanger, Dominique Stoppa-Lyonnet, Marc-Henri Stern

   Pages 25-42

5. An HTRF® Assay for the Protein Kinase ATM

   • Phillip Adams, Jonathan Clark, Simon Hawdon, Jennifer Hill, Andrew Plater

   Pages 43-56

6. ATM Kinase Inhibitors: HTS Cellular Imaging Assay Using Cellomics™ ArrayScan VTI Platform

   • Catherine Bardelle, Joanna Boros

   Pages 57-70

7. Image-Based High Content Screening: Automating the Quantification Process for DNA Damage-Induced Foci

   • Yi Chieh Lim

   Pages 71-84

8. Analyzing ATM Function by Electroporation of Endonucleases and Immunofluorescence Microscopy

   • Keiji Suzuki

   Pages 85-96

9. Quantitative and Dynamic Imaging of ATM Kinase Activity by Bioluminescence Imaging

   • Shyam Nyati, Grant Young, Brian Dale Ross, Alnawaz Rehemtulla

   Pages 97-111

10. Zn(II)–Phos-Tag SDS-PAGE for Separation and Detection of a DNA Damage-Related Signaling Large Phosphoprotein

    • Eiji Kinoshita, Emiko Kinoshita-Kikuta, Tohru Koike

    Pages 113-126

11. Identification of ATM Protein Kinase Phosphorylation Sites by Mass Spectrometry

    • Mark E. Graham, Martin F. Lavin, Sergei V. Kozlov

    Pages 127-144

12. Studies of ATM Kinase Activity Using Engineered ATM Sensitive to ATP Analogues (ATM-AS)

    • Masato Enari, Yuko Matsushima-Hibiya, Makoto Miyazaki, Ryo Otomo

    Pages 145-156

13. Functional Characterization of ATM Kinase Using Acetylation-Specific Antibodies

    • Yingli Sun, Fengxia Du

    Pages 157-162

14. Identification of ATM-Interacting Proteins by Co-immunoprecipitation and Glutathione-S-Transferase (GST) Pull-Down Assays

    • Amanda L. Bain, Janelle L. Harris, Kum Kum Khanna

    Pages 163-181

15. ATM Activation and H2AX Phosphorylation Induced by Genotoxic Agents Assessed by Flow- and Laser Scanning Cytometry

    • Hong Zhao, H. Dorota Halicka, Jorge Garcia, Jiangwei Li, Zbigniew Darzynkiewicz

    Pages 183-196

16. Peptide Immunoaffinity Enrichment with Targeted Mass Spectrometry: Application to Quantification of ATM Kinase Phospho-Signaling

    • Jeffrey R. Whiteaker, Lei Zhao, Regine M. Schoenherr, Jacob J. Kennedy, Richard G. Ivey, Amanda G. Paulovich

    Pages 197-213

17. Mass Spectrometry-Based Proteomics for Quantifying DNA Damage-Induced Phosphorylation

    • Marina E. Borisova, Sebastian A. Wagner, Petra Beli

    Pages 215-227

18. Statistical Analysis of ATM-Dependent Signaling in Quantitative Mass Spectrometry Phosphoproteomics

    • Ashley J. Waardenberg

    Pages 229-244

19. ChIP Technique to Study Protein Dynamics at Defined DNA Double Strand Breaks

    • Jie Wen, Patrick Concannon

    Pages 245-262

20. Studies of the DNA Damage Response by Using the Lac Operator/Repressor (LacO/LacR) Tethering System

    • Rossana Piccinno, Marta Cipinska, Vassilis Roukos

    Pages 263-275

Ataxia–telangiectasia (A-T) is a rare and severe genetic disorder affecting children. A-T is a multisystem disease characterized by progressive neurodegeneration, immunodeficiency and cancer predisposition. This detailed volume explores the ever expanding field of research into the ATM (ataxia-telangiectasia, mutated) gene and the role played by ATM kinase in DNA damage signaling and diverse cellular processes. What follows is a handy desktop reference for both seasoned A-T researchers and postgraduate students, as it demonstrates the breadth of recent developments in A-T studies. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Both classic and cutting-edge techniques are described, including ATM gene mutation detection, assays for radiosensitivity and radioresistant DNA synthesis, new methods to measure ATM kinase activity by imaging microscopy and high content screening as well as proteomics, phosphoproteomics and bioinformatics approaches to decipher ATM-dependent signalling pathways. Additional methods include generation of patient-specific stem cells and approaches to study ATM functions in the nervous system. 

Comprehensive and practical, ATM Kinase: Methods and Protocols aims to ignite and attract the interest of colleagues from diverse fields to A-T research in an effort to bring their expertise and fresh ideas to resolve many A-T puzzles still waiting to be pieced together and to alleviate the suffering of A-T children and their families.

• Ataxia–telangiectasia (A-T)
• Genetic disorder
• DNA damage
• Mutated
• ATM gene

• University of Queensland Centre for Clinical Research (UQCCR), University of Queensland, Herston, Australia

  Sergei V. Kozlov

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