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Protein Kinase C Promotes Notch1 Cleavage Through Both Ligand-Dependent and Hyaluronic Acid-ADAM10-Dependent

Xin Tang1,2,3,4, Yunfei Mu2,3,4, Hongjun Shi2,3,4

  • 1College of Life Sciences, Zhejiang University, Hangzhou 310058, China.

Cells
|August 13, 2026
PubMed

Insights

Protein kinase C (PKC) activates Notch1 signaling in endothelial cells through two pathways: upregulating Notch ligands and promoting hyaluronan synthesis. This reveals the extracellular matrix as a key regulator of Notch1 signal strength.

Area of Science:

  • Endothelial cell biology
  • Molecular signaling pathways
  • Extracellular matrix interactions

Background:

  • Notch1 signaling is crucial for vascular development and endothelial cell fate.
  • Protein kinase C (PKC) is known to activate Notch1, but the precise mechanisms are unclear.

Purpose of the Study:

  • To elucidate the molecular mechanisms by which PKC activates Notch1 signaling in endothelial cells.
  • To identify the role of extracellular matrix components in PKC-mediated Notch1 activation.

Main Methods:

  • Utilized murine endothelial MS1 cells and mouse aortic endothelial cells (MAECs).
  • Investigated the effects of PKC activation on Notch1 ligands (Jag1, Dll4), hyaluronic acid synthase 2 (HAS2), and hyaluronan (HA) synthesis.
  • Employed genetic (knockdown/overexpression) and pharmacological (4-MU) approaches to assess the role of HA.
  • Examined ADAM10 localization and Notch1 phosphorylation.

Main Results:

  • PKC activation upregulates Jag1 and Dll4, creating a positive feedback loop for Notch signaling.
  • PKC enhances HAS2 expression, leading to increased hyaluronan (HA) synthesis, which is essential for Notch1 activation.
  • HA signaling is required for PKC to activate Notch1, and its inhibition downregulates cell surface ADAM10.
  • Direct phosphorylation of Notch1 at S1791 is not required for PKC-mediated activation.

Conclusions:

  • PKC employs a dual pathway involving ligand upregulation and HA signaling to promote Notch1 proteolytic cleavage.
  • The extracellular matrix, specifically HA, acts as a critical regulator of Notch1 signal strength in endothelial cells.
  • These findings uncover a novel mechanism linking PKC, extracellular matrix, and Notch1 signaling in vascular biology.
Keywords:
ADAM10HAS2Jagged1NICDNotch signalingendothelial cellshyaluronic acidprotein kinase C

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