p53 Protocols

1. Front Matter

   Pages i-xi

   PDF

2. Adenovirus Expressing p53

   • Matthew Holmes, Elizabeth Rosenberg, Kristoffer Valerie

   Pages 1-16

3. Purification of Recombinant p53 from Sf9 Insect Cells

   • Xiu Zhu Sun, John Nguyen, Jamil Momand

   Pages 17-28

4. Generation and Characterization of p53 Mutant Mice

   • Melissa L. Dumble, Lawrence A. Donehower, Xiongbin Lu

   Pages 29-49

5. Identification of p53 Target Genes by Fluorescent Differential Display

   • Susanne Stein, Yong-Jig Cho, Roger S. Jackson II, Peng Liang

   Pages 51-63

6. p53-Induced Gene Expression Analysis

   • Joanna K. Sax, Wafik S. El-Deiry

   Pages 65-71

7. Real-Time Polymerase Chain Reaction Quantitation of Relative Expression of Genes Modulated by p53 Using SYBR® Green I

   • Katherine E. Roth Stagliano, Evie Carchman, Sumitra Deb

   Pages 73-91

8. Transactivation and Transrepression Studies with p53

   • Mariano J. Scian, Rebecca Frum, Swati Palit Deb, Sumitra Deb

   Pages 93-110

9. Methods to Study p53-Repressed Promoters

   • Patrick Dumont, Anthony Della Pietra, Maureen E. Murphy

   Pages 111-120

10. Interaction of p53 with Cellular Proteins

    • David M. Keller, Shelya X. Zeng, Hua Lu

    Pages 121-133

11. Interaction of p53 with the Adenovirus E1B-55 kDa Protein

    • Judith Roth, Matthias Dobbelstein

    Pages 135-149

12. In Vivo Footprinting and DNA Affinity Chromatography for Analysis of p53 DNA Binding Ability

    • Maria Patricia Molina, Christine Cain, Jill Bargonetti

    Pages 151-170

13. Signaling to p53

    • Ashley L. Craig, Susan E. Bray, Lee E. Finlan, Neil M. Kernohan, Ted R. Hupp

    Pages 171-202

14. Detection of Mitochondrial Localization of p53

    • Motohiro Mihara, Ute M. Moll

    Pages 203-209

15. Analysis of Nuclear and Cytoplasmic Degradation of p53 in Cells after Stress

    • Troy W. Joseph, Ute M. Moll

    Pages 211-217

16. Mutational Analysis of p53 in Human Tumors

    • Susan Erster, Neda Slade, Ute M. Moll

    Pages 219-230

17. Mutational Analysis of p53 in Human Tumors

    • Neda Slade, Ute M. Moll

    Pages 231-243

18. Flow Cytometric Analysis of p53-Induced Apoptosis

    • Michael Berger, Ygal Haupt

    Pages 245-256

19. Flow Cytometric Analysis of MDM2-Mediated Growth Arrest

    • Rebecca Frum, Swati Palit Deb

    Pages 257-267

20. Back Matter

    Pages 269-280

    PDF

Since the discovery of p53 as a tumor suppressor, numerous methods have evolved to reveal the unique structural features and biochemical functions of this protein. Several unique properties of p53 posed a challenge to understa- ing its normal function in the initial phase of its research. The low levels of p53 in normal cells, its stabilization under situations of genotoxic stress, induction of growth arrest, and apoptosis with stabilization of the protein, obstructed the visibility of its normal, unmutated function. The property of p53 that can sense a promoter and transactivate or inhibit is still not well understood. It is still not known whether it is the absence of the protein that causes tumorigenesis, or if its mutants have a dominant role in inducing cancer. p53 Protocols comprises eighteen chapters for the study of the diverse properties of p53 and related proteins. The methods included are invaluable for delineating the function of other proteins that may function as tumor suppr- sors or growth suppressors. The chapters are not presented in any schematic order, for the importance and diversity of the functions of p53 make it imp- sible to organize them suitably. We have made a sincere effort to collect the methods most useful to those investigators working on tumor suppressors or growth suppressors. The purpose of p53 Protocols is not only to provide investigators with methods to analyze similar biochemical functions, but also to familiarize them with the associated problems that arose during the course of investigations.

• Medical College of Virginia, Virginia Commonwealth University, Richmond

  Sumitra Deb, Swati Palit Deb

Policies and ethics