An essential cytoplasmic domain for the Golgi localization of coiled-coil proteins with a COOH-terminal membrane anchor.
Giantin is a resident Golgi protein that has an extremely long cytoplasmic domain (about 370 kDa) and is anchored to the Golgi membrane by the COOH-terminal membrane-anchoring domain (CMD) with no luminal extension. We examined the essential domain of giantin required for Golgi localization by mutational analysis. The Golgi localization of giantin was not affected by the deletion of its CMD or by substitution with the CMD of syntaxin-2, a plasma membrane protein. The giantin CMD fused to the cytoplasmic domain of syntaxin-2 could not retain the chimera in the Golgi apparatus. Sequential deletion analysis showed that the COOH-terminal sequence (positions 3059--3161) adjacent to the CMD was the essential domain required for the Golgi localization of giantin. We also examined two other Golgi-resident proteins, golgin-84 and syntaxin-5, with a similar membrane topology as giantin. It was confirmed that the cytoplasmic domain of about 100 residues adjacent to the CMD was required for their Golgi localization. Taken together, these results suggest that the COOH-terminally anchored Golgi proteins with long cytoplasmic extensions have the Golgi localization signal(s) in the cytoplasmic sequence adjacent to the CMD. This is in contrast to previous observations that a transmembrane domain is required for Golgi localization by other Golgi proteins transported from the endoplasmic reticulum.
Misumi
Y
Y
Department of Biochemistry, Fukuoka University School of Medicine, Jonan-ku, Fukuoka 814-0180, Japan.
Sohda
M
M
Tashiro
A
A
Sato
H
H
Ikehara
Y
Y
D000818:Animals; D001324:Autoantigens / Q000737:chemistry; D019556:COS Cells; D003593:Cytoplasm / Q000737:chemistry*; D004721:Endoplasmic Reticulum / Q000737:chemistry; D006056:Golgi Apparatus / Q000737:chemistry*; D000077614:Golgi Matrix Proteins; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry*; D050765:Qa-SNARE Proteins
D001324:Autoantigens; C098772:GOLGA4 protein, human; D000077614:Golgi Matrix Proteins; D008565:Membrane Proteins; D050765:Qa-SNARE Proteins; C087574:macrogolgin