Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Cyclin a-CDK phosphorylation regulates MDM2 protein interactions.

T Zhang1, C Prives

  • 1Department of Biological Sciences, Columbia University, New York, New York 10027, USA.

The Journal of Biological Chemistry
|May 22, 2001
PubMed
Summary

MDM2 protein phosphorylation by cyclin A-CDK2 at Thr-216 weakens its interaction with p53 and alters its binding to p19ARF. This modification, prevalent during S phase, affects MDM2 antibody recognition.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Correction: Energy-dependent nucleolar localization of p53 in vitro requires two discrete regions within the p53 carboxyl terminus.

Oncogene·2018
Same author

Pro-proliferative FoxM1 is a target of p53-mediated repression.

Oncogene·2009
Same author

p53 responsive elements in human retrotransposons.

Oncogene·2009
Same author

Are interactions with p63 and p73 involved in mutant p53 gain of oncogenic function?

Oncogene·2007
Same author

Energy-dependent nucleolar localization of p53 in vitro requires two discrete regions within the p53 carboxyl terminus.

Oncogene·2007
Same author

Transcriptional regulation by p53 and p73.

Cold Spring Harbor symposia on quantitative biology·2006

Area of Science:

  • Molecular Biology
  • Cell Cycle Regulation
  • Oncogenesis

Background:

  • MDM2 protein regulates the tumor suppressor p53.
  • Post-translational modifications of MDM2 influence its cellular functions.
  • Cyclin-dependent kinases (CDKs) are key regulators of the cell cycle.

Purpose of the Study:

  • To investigate the role of MDM2 phosphorylation in regulating its interaction with p53 and p19ARF.
  • To identify the specific kinase responsible for MDM2 phosphorylation and the site of modification.
  • To correlate MDM2 phosphorylation with cell cycle progression.

Main Methods:

  • In vitro kinase assays using murine MDM2 and various cyclin-CDK complexes.
  • Peptide inhibition assays to identify cyclin recognition motifs.

Related Experiment Videos

  • Two-dimensional phosphopeptide mapping and mutational analysis to pinpoint phosphorylation sites.
  • Western blot analysis using a specific monoclonal antibody (SMP14) to detect phosphorylated MDM2 in cell extracts.

    • Main Results:

    • MDM2 is efficiently phosphorylated by cyclin A-CDK2 and cyclin A-CDK1, but not by other cyclin-containing complexes.
    • Phosphorylation occurs at Thr-216, weakening MDM2 binding to p53 and enhancing binding to p19ARF.
    • Phosphorylation at Thr-216 prevents recognition by the MDM2-specific antibody SMP14.
    • In vivo, Thr-216 phosphorylation is most prominent at the start of S phase, coinciding with cyclin A expression.

    Conclusions:

    • Cyclin A-CDK2-mediated phosphorylation of MDM2 at Thr-216 is a key regulatory mechanism.
    • This phosphorylation event modulates the interaction of MDM2 with its binding partners p53 and p19ARF.
    • MDM2 phosphorylation at Thr-216 serves as a cell cycle-dependent marker, detectable during S phase.