GSK3 Inhibits Macropinocytosis and Lysosomal Activity through the Wnt Destruction Complex Machinery

Cell Rep. 2020 Jul 28;32(4):107973. doi: 10.1016/j.celrep.2020.107973.

Abstract

Canonical Wnt signaling is emerging as a major regulator of endocytosis. Here, we report that Wnt-induced macropinocytosis is regulated through glycogen synthase kinase 3 (GSK3) and the β-catenin destruction complex. We find that mutation of Axin1, a tumor suppressor and component of the destruction complex, results in the activation of macropinocytosis. Surprisingly, inhibition of GSK3 by lithium chloride (LiCl), CHIR99021, or dominant-negative GSK3 triggers macropinocytosis. GSK3 inhibition causes a rapid increase in acidic endolysosomes that is independent of new protein synthesis. GSK3 inhibition or Axin1 mutation increases lysosomal activity, which can be followed with tracers of active cathepsin D, β-glucosidase, and ovalbumin degradation. Microinjection of LiCl into the blastula cavity of Xenopus embryos causes a striking increase in dextran macropinocytosis. The effects of GSK3 inhibition on protein degradation in endolysosomes are blocked by the macropinocytosis inhibitors EIPA or IPA-3, suggesting that increases in membrane trafficking drive lysosomal activity.

Keywords: Axin1; Pak1; cathepsin D; colorectal carcinoma; hepatocellular carcinoma; lysosome; membrane trafficking; multivesicular bodies; nutrient acquisition; β-glucosidase.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Axin Protein / metabolism*
  • Cell Line, Tumor
  • Endocytosis / physiology
  • Endosomes / metabolism
  • Glycogen Synthase Kinase 3 / metabolism*
  • Glycogen Synthase Kinase 3 / physiology
  • Lysosomes / metabolism
  • Phosphorylation
  • Pinocytosis / physiology*
  • Wnt Proteins / metabolism
  • Wnt Signaling Pathway / physiology
  • Xenopus Proteins / metabolism*
  • Xenopus Proteins / physiology
  • Xenopus laevis
  • beta Catenin / metabolism

Substances

  • Axin Protein
  • Wnt Proteins
  • Xenopus Proteins
  • axin1 protein, Xenopus
  • beta Catenin
  • GSK3B protein, Xenopus
  • Glycogen Synthase Kinase 3