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

Regulation of Id3 cell cycle function by Cdk-2-dependent phosphorylation

R W Deed1, E Hara, G T Atherton

  • 1CRC Department of Gene Regulation, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester, United Kingdom.

Molecular and Cellular Biology
|December 31, 1997
PubMed
Summary

Cell cycle-regulated phosphorylation of Id3 protein by Cdk2 alters its function in regulating gene expression and cell proliferation. This phosphorylation acts as a switch, impacting cell cycle progression and the integration of Id and bHLH protein functions.

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

Glycine synthesis from nitrate and glyoxylate mediated by ferroan brucite: An integrated pathway for prebiotic amine synthesis.

Proceedings of the National Academy of Sciences of the United States of America·2024
Same author

Reversion-inducing cysteine-rich protein with Kazal motifs interferes with epidermal growth factor receptor signaling.

Oncogene·2010
Same author

Epigenetic abnormalities in cutaneous squamous cell carcinomas: frequent inactivation of the RB1/p16 and p53 pathways.

The British journal of dermatology·2006
Same author

Site-specific DNA methylation by a complex of PU.1 and Dnmt3a/b.

Oncogene·2005
Same author

Cancer immunotherapy by fusions of dendritic and tumour cells and rh-IL-12.

European journal of clinical investigation·2005
Same author

Detection of Staphylococcus aureus by 16S rRNA directed in situ hybridisation in a patient with a brain abscess caused by small colony variants.

Journal of neurology, neurosurgery, and psychiatry·2003

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Basic helix-loop-helix (bHLH) transcription factors regulate cell differentiation and cell cycle arrest.
  • Id proteins, a family of HLH proteins, inhibit bHLH transcription factor activity by sequestering them into inactive heterodimers.
  • Phosphorylation of Id proteins by cyclin-dependent kinases (Cdks) has been implicated in cell cycle regulation.

Purpose of the Study:

  • To investigate the role of cell-cycle-regulated phosphorylation of Id3 protein in modulating its function.
  • To determine how Id3 phosphorylation affects its interaction with bHLH proteins and its influence on gene expression and cell cycle progression.
  • To elucidate the mechanism by which Cdk2-dependent phosphorylation integrates Id and bHLH functions in cell proliferation and differentiation.

Related Experiment Videos

Main Methods:

  • Site-directed mutagenesis to create Id3 phosphorylation mimic (Asp5) and non-phosphorylatable (Ala5) mutants.
  • In vitro assays to assess the ability of Id3 and its mutants to abrogate bHLH homo- and heterodimer complex formation.
  • In vivo reporter gene assays to evaluate the effect of Id3 mutants on E-box-dependent gene expression.
  • Transfection studies in fibroblasts to assess the impact of Id3 mutants on cell cycle S phase entry.

Main Results:

  • Cell-cycle-regulated phosphorylation of Id3 alters its specificity in abrogating bHLH complex formation and E-box-dependent reporter gene function.
  • An Id3 Asp5 mutant (mimicking phosphorylation) failed to promote cell cycle S phase entry.
  • An Id3 Ala5 mutant (ablating phosphorylation) showed enhanced activity in promoting S phase entry compared to wild-type Id3.
  • Cdk2-dependent phosphorylation of Id3 acts as a switch during late G1-to-S phase, nullifying an early G1 function and modulating bHLH target specificity.

Conclusions:

  • Cdk2-dependent phosphorylation of Id3 is a critical regulatory mechanism controlling its function in cell cycle progression.
  • The ability of Id3 to promote S phase entry is not solely dependent on its modulation of bHLH heterodimer-dependent gene expression.
  • A biological mechanism exists for integrating Cdk2 and Id-bHLH functions to coordinate cell proliferation and differentiation.