In situ cleavage of the acidic domain from the p115 tether inhibits exocytic transport.
Golgins are coiled-coil proteins involved in Golgi architecture and function. A complex of golgins (p115, GM130 and giantin), together with the rab1 guanosine triphosphatase and cis Golgi SNAREs, helps to mediate fusion processes at the entry face of the Golgi apparatus. The C-terminal acidic domain of p115 binds specifically to GM130 and giantin. However, deletion of this domain in vivo appears to have no effect on exocytic transport when using an RNA interference depletion/rescue approach (Puthenveedu MA, Linstedt AD. Gene replacement reveals that p115/SNARE interactions are essential for Golgi biogenesis. Proc Natl Acad Sci U S A 2004;101:1253-1256). In this study, we have used a different approach introducing a tobacco etch virus (tev) protease cleavage site into p115 so that the C-terminal domain can be rapidly and specifically released in vivo by microinjection of the tev protease. The results show that cleavage inhibits exocytic transport to the cell surface.
Satoh
Ayano
A
Department of Cell Biology, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06520-8002, USA.
Warren
Graham
G
D001324:Autoantigens / Q000378:metabolism; D001665:Binding Sites; D015153:Blotting, Western; D020755:Coat Protein Complex I / Q000378:metabolism; D010450:Endopeptidases / Q000378:metabolism / Q000494:pharmacology; D005089:Exocytosis; D006056:Golgi Apparatus / Q000378:metabolism*; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D047468:Immunoprecipitation; D008561:Membrane Fusion; D008565:Membrane Proteins / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology; D011485:Protein Binding; D021381:Protein Transport; D033921:Vesicular Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
D001324:Autoantigens; D020755:Coat Protein Complex I; D000077614:Golgi Matrix Proteins; C097741:Golgin subfamily A member 2; D008565:Membrane Proteins; D010446:Peptide Fragments; D033921:Vesicular Transport Proteins; C087574:macrogolgin; C091580:vesicular transport factor p115; D010450:Endopeptidases; C099488:TEV protease