Differential stability of beta-catenin along the animal-vegetal axis of the sea urchin embryo mediated by dishevelled

Development. 2004 Jun;131(12):2947-56. doi: 10.1242/dev.01152. Epub 2004 May 19.

Abstract

beta-Catenin has a central role in the early axial patterning of metazoan embryos. In the sea urchin, beta-catenin accumulates in the nuclei of vegetal blastomeres and controls endomesoderm specification. Here, we use in-vivo measurements of the half-life of fluorescently tagged beta-catenin in specific blastomeres to demonstrate a gradient in beta-catenin stability along the animal-vegetal axis during early cleavage. This gradient is dependent on GSK3beta-mediated phosphorylation of beta-catenin. Calculations show that the difference in beta-catenin half-life at the animal and vegetal poles of the early embryo is sufficient to produce a difference of more than 100-fold in levels of the protein in less than 2 hours. We show that dishevelled (Dsh), a key signaling protein, is required for the stabilization of beta-catenin in vegetal cells and provide evidence that Dsh undergoes a local activation in the vegetal region of the embryo. Finally, we report that GFP-tagged Dsh is targeted specifically to the vegetal cortex of the fertilized egg. During cleavage, Dsh-GFP is partitioned predominantly into vegetal blastomeres. An extensive mutational analysis of Dsh identifies several regions of the protein that are required for vegetal cortical targeting, including a phospholipid-binding motif near the N-terminus.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Animals
  • Body Patterning
  • Cloning, Molecular
  • Cytoskeletal Proteins / metabolism*
  • Dishevelled Proteins
  • Embryo, Nonmammalian / cytology
  • Embryo, Nonmammalian / physiology
  • Glycogen Synthase Kinase 3 / genetics
  • Glycogen Synthase Kinase 3 / metabolism
  • Glycogen Synthase Kinase 3 beta
  • Kinetics
  • Phosphoproteins / genetics*
  • Phosphoproteins / metabolism
  • Recombinant Proteins / metabolism
  • Sea Urchins / embryology*
  • Signal Transduction / physiology
  • Trans-Activators / metabolism*
  • beta Catenin

Substances

  • Adaptor Proteins, Signal Transducing
  • Cytoskeletal Proteins
  • Dishevelled Proteins
  • Phosphoproteins
  • Recombinant Proteins
  • Trans-Activators
  • beta Catenin
  • Glycogen Synthase Kinase 3 beta
  • Glycogen Synthase Kinase 3