Direct binding of the Kex2p cytosolic tail to the VHS domain of yeast Gga2p facilitates TGN to prevacuolar compartment transport and is regulated by phosphorylation
| العنوان: | Direct binding of the Kex2p cytosolic tail to the VHS domain of yeast Gga2p facilitates TGN to prevacuolar compartment transport and is regulated by phosphorylation |
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| المؤلفون: | Mithu De, Mohamed E. Abazeed, Robert S. Fuller |
| المصدر: | Molecular Biology of the Cell. 24:495-509 |
| بيانات النشر: | American Society for Cell Biology (ASCB), 2013. |
| سنة النشر: | 2013 |
| مصطلحات موضوعية: | 0303 health sciences, Endosome, Saccharomyces cerevisiae, Cell Biology, Plasma protein binding, Biology, biology.organism_classification, Cell biology, Transport protein, 03 medical and health sciences, A-site, 0302 clinical medicine, Biochemistry, Phosphorylation, Signal transduction, Binding site, Molecular Biology, 030217 neurology & neurosurgery, 030304 developmental biology |
| الوصف: | Human Golgi-localized, γ-ear–containing, ADP-ribosylation factor–binding proteins (Ggas) bind directly to acidic dileucine sorting motifs in the cytosolic tails (C-tails) of intracellular receptors. Despite evidence for a role in recruiting ubiquitinated cargo, it remains unclear whether yeast Ggas also function by binding peptide-sorting signals directly. Two-hybrid analysis shows that the Gga1p and Gga2p Vps27, Hrs, Stam (VHS) domains both bind a site in the Kex2p C-tail and that the Gga2p VHS domain binds a site in the Vps10p C-tail. Binding requires deletion of an apparently autoinhibitory sequence in the Gga2p hinge. Ser780in the Kex2p C-tail is crucial for binding: an Ala substitution blocks but an Asp substitution permits binding. Biochemical assays using purified Gga2p VHS–GGA and TOM1 (GAT) and glutathione S-transferase–Kex2p C-tail fusions show that Gga2p binds directly to the Kex2p C-tail, with relative affinities Asp780> Ser780> Ala780. Affinity-purified antibody against a peptide containing phospho-Ser780recognizes wild-type Kex2p but not S780A Kex2p, showing that Ser780is phosphorylated in vivo; phosphorylation of Ser780is up-regulated by cell wall–damaging drugs. Finally, mutation of Ser780alters trafficking of Kex2p both in vivo and in cell-free trans-Golgi network (TGN)–prevacuolar compartment (PVC) transport. Thus yeast Gga adaptors facilitate TGN–PVC transport by direct binding of noncanonical phosphoregulated Gga-binding sites in cargo molecules. |
| تدمد: | 1939-4586 1059-1524 |
| DOI: | 10.1091/mbc.e12-04-0322 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::ac46773ef59afcb4692df9e8b3cd8d31 https://doi.org/10.1091/mbc.e12-04-0322 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi...........ac46773ef59afcb4692df9e8b3cd8d31 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19394586 10591524 |
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| DOI: | 10.1091/mbc.e12-04-0322 |