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Activation of the p21(CIP1/WAF1) promoter by bone morphogenetic protein-2 in mouse B lineage cells

K Yamato1, S Hashimoto, T Imamura

  • 1Section of Molecular Cellular Oncology and Microbiology, Division of Oral Health Science, Graduate School, Tokyo Medical and Dental University, Tokyo 113-8549, Japan.

Oncogene
|July 24, 2001
PubMed

Insights

Bone Morphogenetic Proteins (BMPs) like BMP-2 inhibit cell growth by activating the p21 gene. This study identifies a specific DNA region where Smad proteins bind, mediating BMP-2

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • Bone Morphogenetic Proteins (BMPs) are known to inhibit cell growth.
  • Previous research indicated BMP-2 induces p21(CIP1/WAF1) expression, inhibiting HS-72 mouse hybridoma cell growth.

Purpose of the Study:

  • To elucidate the mechanism by which BMP-2 activates the mouse p21(CIP1/WAF1) promoter in HS-72 cells.
  • To identify the specific DNA region and protein factors involved in BMP-2-mediated p21(CIP1/WAF1) transcriptional activation.

Main Methods:

  • Reporter assays to assess promoter activity.
  • Site-directed mutagenesis to identify responsive promoter regions.
  • Electrophoretic mobility shift assays (EMSAs) to detect protein-DNA interactions.
  • Western blotting to detect protein expression and phosphorylation.

Main Results:

  • BMP-2 activated the mouse p21(CIP1/WAF1) promoter in HS-72 cells.
  • A conserved 29-base pair region (-1928/-1900) of the p21 promoter responded to BMP-2, Smad1, Smad4, and activated BMP type I receptors.
  • This 29-bp region specifically bound Smad4 and phosphorylated Smad1 in BMP-2-stimulated cells.

Conclusions:

  • BMP-2 activates p21(CIP1/WAF1) transcription in HS-72 cells.
  • The mechanism involves the binding of Smad4 and Smad1 to a conserved 29-bp region of the p21 promoter.
  • This Smad-binding element is crucial for BMP-2-induced growth inhibition via p21 activation.

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