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Related Experiment Videos

NPAT expression is regulated by E2F and is essential for cell cycle progression.

Guang Gao1, Adrian P Bracken, Karina Burkard

  • 1Department of Biomedical Genetics, University of Rochester, Rochester, New York 14642, USA.

Molecular and Cellular Biology
|April 1, 2003
PubMed
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Nuclear protein AT (NPAT) is a crucial E2F target gene essential for cell cycle progression. NPAT links E2F transcription factors to the activation of S-phase-specific histone gene transcription, facilitating cell division.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • Nuclear protein AT (NPAT) is a known substrate of cyclin E-Cdk2 kinase.
  • NPAT is implicated in regulating histone gene transcription during the G1/S-phase transition and S-phase entry in mammalian cells.

Purpose of the Study:

  • To investigate the regulation of NPAT gene expression by E2F transcription factors.
  • To elucidate the role of NPAT in cell cycle progression and histone gene transcription.

Main Methods:

  • Analysis of NPAT promoter activity in response to E2F activation.
  • In vivo chromatin immunoprecipitation (ChIP) to detect E2F binding to the NPAT promoter.
  • Quantitative real-time PCR (qRT-PCR) to measure NPAT mRNA levels.
  • Small interfering RNA (siRNA) to inhibit NPAT expression.

Related Experiment Videos

  • Cell cycle analysis.

    • Main Results:

    • NPAT transcription is upregulated at the G1/S-phase boundary in growth-stimulated cells.
    • Endogenous E2F proteins bind to the NPAT promoter in vivo, and E2F1 overexpression stimulates NPAT mRNA expression.
    • E2F binding sites within the NPAT promoter are essential for its activation during the G1/S-phase transition.
    • NPAT expression accelerates S-phase entry.
    • Inhibition of NPAT expression by siRNA impedes cell cycle progression and histone gene expression.

    Conclusions:

    • NPAT is a key E2F target gene required for mammalian cell cycle progression.
    • NPAT acts as a molecular link between E2F transcription factors and the activation of S-phase-specific histone gene transcription.