Localization of glypican-4 in different membrane microdomains is involved in the regulation of Wnt signaling.
Glypicans are members of the heparan sulfate proteoglycans (HSPGs) and are involved in various growth factor signaling mechanisms. Although HSPGs affect the β-catenin-dependent and -independent pathways of Wnt signaling, how they regulate distinct Wnt pathways is not clear. It has been suggested that the β-catenin-dependent pathway is initiated through receptor endocytosis in lipid raft microdomains and the independent pathway is activated through receptor endocytosis in non-lipid raft microdomains. Here, evidence is presented that glypican-4 (GPC4) is localized to both membrane microdomains and that the localization affects its ability to regulate distinct Wnt pathways. GPC4 bound to Wnt3a and Wnt5a, which activate the β-catenin-dependent and -independent pathways, respectively, and colocalized with Wnts on the cell surface. LRP6, one of Wnt3a coreceptors, was present in lipid raft microdomains, whereas Ror2, one of Wnt5a coreceptors, was localized to non-lipid raft microdomains. Expression of GPC4 enhanced the Wnt3a-dependent β-catenin pathway and the Wnt5a-dependent β-catenin-independent pathway, and knockdown of GPC4 suppressed both pathways. A GPC4 mutant that was localized to only non-lipid raft microdomains inhibited the β-catenin-dependent pathway but enhanced the β-catenin-independent pathway. These results suggest that GPC4 concentrates Wnt3a and Wnt5a to the vicinity of their specific receptors in different membrane microdomains, thereby regulating distinct Wnt signaling.
Department of Molecular Biology and Biochemistry, Graduate School of Medicine, Faculty of Medicine, Osaka University, Suita 565-0871,  Japan.
Hiroshi
H
Sakane
Hideki
H
Yamamoto
Shinji
S
Matsumoto
Akira
A
Sato
Akira
A
Kikuchi
D002460:Cell Line; D066246:ErbB Receptors / Q000737:chemistry; D053673:Glypicans / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D021962:Membrane Microdomains / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D060490:Receptors, Wnt / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D053668:Syndecan-1 / Q000737:chemistry; D051153:Wnt Proteins / Q000494:pharmacology*; D060449:Wnt Signaling Pathway; D060509:Wnt3A Protein / Q000378:metabolism / Q000494:pharmacology; D051176:beta Catenin / Q000378:metabolism
C501126:DKK1 protein, human; C492044:GPC4 protein, human; D053673:Glypicans; D036341:Intercellular Signaling Peptides and Proteins; D060490:Receptors, Wnt; D011993:Recombinant Fusion Proteins; D053668:Syndecan-1; D051153:Wnt Proteins; D060509:Wnt3A Protein; D051176:beta Catenin; D066246:ErbB Receptors