E-cadherin transport from the trans-Golgi network in tubulovesicular carriers is selectively regulated by golgin-97.
E-cadherin is a cell-cell adhesion protein that is trafficked and delivered to the basolateral cell surface. Membrane-bound carriers for the post-Golgi exocytosis of E-cadherin have not been characterized. Green fluorescent protein (GFP)-tagged E-cadherin (Ecad-GFP) is transported from the trans-Golgi network (TGN) to the recycling endosome on its way to the cell surface in tubulovesicular carriers that resemble TGN tubules labeled by members of the golgin family of tethering proteins. Here, we examine the association of golgins with tubular carriers containing E-cadherin as cargo. Fluorescent GRIP domains from golgin proteins replicate the membrane binding of the full-length proteins and were coexpressed with Ecad-GFP. The GRIP domains of p230/golgin-245 and golgin-97 had overlapping but nonidentical distributions on the TGN; both domains were on TGN-derived tubules but only the golgin-97 GRIP domain coincided with Ecad-GFP tubules in live cells. When the Arl1-binding endogenous golgins, p230/golgin-245 and golgin-97 were displaced from Golgi membranes by overexpression of the p230 GRIP domain, trafficking of Ecad-GFP was inhibited. siRNA knockdown of golgin-97 also inhibited trafficking of Ecad-GFP. Thus, the GRIP domains of p230/golgin-245 and golgin-97 bind discriminately to distinct membrane subdomains of the TGN. Golgin-97 is identified as a selective and essential component of the tubulovesicular carriers transporting E-cadherin out of the TGN.
Lock
John G
JG
Institute for Molecular Bioscience, The University of Queensland, Brisbane, Queensland 4072, Australia.
Hammond
Luke A
LA
Houghton
Fiona
F
Gleeson
Paul A
PA
Stow
Jennifer L
JL
D001324:Autoantigens / Q000502:physiology*; D015820:Cadherins / Q000378:metabolism*; D002352:Carrier Proteins / Q000502:physiology*; D017930:Genes, Reporter; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000502:physiology; D008870:Microtubules / Q000378:metabolism; D021381:Protein Transport / Q000502:physiology; D021601:trans-Golgi Network / Q000378:metabolism*
D001324:Autoantigens; D015820:Cadherins; D002352:Carrier Proteins; C098772:GOLGA4 protein, human; D000077614:Golgi Matrix Proteins; C450721:Golgi complex autoantigen, 97-kDa; D008565:Membrane Proteins