title: A role for the lumenal domain in Golgi localization of the Saccharomyces cerevisiae guanosine diphosphatase.
abstract: Integral membrane proteins (IMPs) contain localization signals necessary for targeting to their resident subcellular compartments. To define signals that mediate localization to the Golgi complex, we have analyzed a resident IMP of the Saccharomyces cerevisiae Golgi complex, guanosine diphosphatase (GDPase). GDPase, which is necessary for Golgi-specific glycosylation reactions, is a type II IMP with a short amino-terminal cytoplasmic domain, a single transmembrane domain (TMD), and a large catalytic lumenal domain. Regions specifying Golgi localization were identified by analyzing recombinant proteins either lacking GDPase domains or containing corresponding domains from type II vacuolar IMPs. Neither deletion nor substitution of the GDPase cytoplasmic domain perturbed Golgi localization. Exchanging the GDPase TMD with vacuolar protein TMDs only marginally affected Golgi localization. Replacement of the lumenal domain resulted in mislocalization of the chimeric protein from the Golgi to the vacuole, but a similar substitution leaving 34 amino acids of the GDPase lumenal domain intact was properly localized. These results identify a major Golgi localization determinant in the membrane-adjacent lumenal region (stem) of GDPase. Although necessary, the stem domain is not sufficient to mediate localization; in addition, a membrane-anchoring domain and either the cytoplasmic or full-length lumenal domain must be present to maintain Golgi residence. The importance of lumenal domain sequences in GDPase Golgi localization and the requirement for multiple hydrophilic protein domains support a model for Golgi localization invoking protein-protein interactions rather than interactions between the TMD and the lipid bilayer.
authors.affiliation: Department of Biological Chemistry, School of Medicine, University of California, Los Angeles, Los Angeles, California 90095, USA.
authors.forename: J J
authors.initials: JJ
authors.lastname: Vowels
authors.forename: G S
authors.initials: GS
authors.lastname: Payne
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D003593:Cytoplasm / Q000378:metabolism; D006056:Golgi Apparatus / Q000201:enzymology* / Q000502:physiology; D008565:Membrane Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D011755:Pyrophosphatases / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
chemical_list: D008565:Membrane Proteins; D011755:Pyrophosphatases; C010357:guanosine-diphosphatase