Proteomic Analysis of Wnt-Dependent Dishevelled-Based Supermolecular Complexes

摘要: Wnt signaling is critical and indispensable for numerous cellular pathways including embryonic development adult tissue homeostasis such as determination, proliferation, migration differentiation (Clevers, 2006; Glass Karsenty, Nusse, 2005; Wang Wynshaw-Boris, 2004). There are three independent branches of the cascade, canonical beta-catenin/Lef-Tcf-sensitive transcriptional response, planar cell polarity (PCP) Wnt5a/Frizzled-2/Ca2+/cGMP response (He et al., 2004; Katanaev Liu 2001; Malbon, Wallingford Habas, Willert 2003). Dishevelleds (Dvls) essential components in major pathways. Dvls function scaffold protein bridging receptors distinct downstream components. Through formation dynamic multiprotein complexes, signal transduced to inside cells. In Wnt/beta-catenin pathway, Wnts bind members Frizzled (Fz) family G protein-coupled co-receptor low density lipoprotein receptor related LRP5 LRP6 (LRP5/6). Binding Wnt3a these facilitates a variety intracellular events. Phosphorylation LRP5/6 triggers interaction Fz-LRP5/6 complex with adenomatous polyposis coli (APC), Dishevelled (Dsh/Dvl), Axin, glycogen synthase kinase-3β฀(GSK3β) casein kinase 1α฀(Hart, 1998; Kishida 1998). Scaffold Dvl destruction complexes (Hart, 1998 ; 2005). As results, stabilized β-catenin, translocates nuclear enhances Lef/Tcf-sensitive transcription (Angers & Moon, 2009; Clevers, van Amerongen 2009). Fly Dsh isoforms mammals (Dvl1-3) all share several prominent, highlyconserved domains: homology domain called DIX; conserved sequence element postsynaptic PSD-95, Discs-large, ZO-1, termed PDZ; Disheveled, Egl-10, Pleckstrin domain, DEP (Wharton, addition, there regions harboring basic amino acids residues prolinerich putative Src 3 (SH3) binding (Penton 2002). Dvl3 knockout mouse lethal, whereas Dvl1 or Dvl2 viable (Hamblet 2002; Etheridge 2008; Lijam 1997). The not truly “redundant” respect Lee 2008), although functions each isoform fully resolved. Previous studies reveal localization totipotent F9 embryonal