| First Author | Yokoyama N | Year | 2010 |
| Journal | J Cell Sci | Volume | 123 |
| Issue | Pt 21 | Pages | 3693-702 |
| PubMed ID | 20940260 | Mgi Jnum | J:182922 |
| Mgi Id | MGI:5317085 | Doi | 10.1242/jcs.075275 |
| Citation | Yokoyama N, et al. (2010) Wnt-dependent assembly of supermolecular Dishevelled-3-based complexes. J Cell Sci 123(Pt 21):3693-702 |
| abstractText | Dishevelled-3 (Dvl3) is a multivalent scaffold protein that is essential to Wnt signaling during development. Although Dvl-based punctae have been visualized by fluorescence microscopy; the physical nature and dynamic character of the such complexes are enigmatic. We use steric-exclusion chromatography, affinity pull-downs, proteomics and fluorescence correlation microscopy to characterize supermolecular Dvl3-based complexes of totipotent mouse F9 cells. The molecular mass of the complexes ranges from that of homodimeric Dvl3 to well-defined peaks harboring supermolecular complexes of 0.4 to 2.0 MDa. Addition of Wnt3a stimulates the formation of Dvl3-based complexes of greater molecular mass within 30 minutes. The presence of DKK1 and knockdown of Dishevelled proteins block formation of the 2 MDa Dvl3-based complexes and also block Wnt3a stimulation of the canonical pathway. Fluorescent correlation microscopy identified supermolecular Dvl3-based complexes with a molecular mass >30 MDa in live cells; these complexes were provoked to form structures with even greater molecular mass by Wnt3a. We establish for the first time the physical and functional nature of very large, supermolecular Dvl3-based complexes. |
