title: COG-7-deficient Human Fibroblasts Exhibit Altered Recycling of Golgi Proteins.
abstract: Recently, we reported that two siblings presenting with the clinical syndrome congenital disorders of glycosylation (CDG) have mutations in the gene encoding Cog7p, a member of the conserved oligomeric Golgi (COG) complex. In this study, we analyzed the localization and trafficking of multiple Golgi proteins in patient fibroblasts under a variety of conditions. Although the immunofluorescent staining pattern of several Golgi proteins was indistinguishable from normal, the staining of endoplasmic reticulum (ER)-Golgi intermediate compartment (ERGIC)-53 and the vesicular-soluble N-ethylmaleimide-sensitive factor attachment protein receptors GS15 and GS28 was abnormal, and the steady-state level of GS15 was greatly decreased. Retrograde transport of multiple Golgi proteins to the ER in patient fibroblasts via brefeldin A-induced tubules was significantly slower than occurs in normal fibroblasts, whereas anterograde protein trafficking was much less affected. After prolonged treatment with brefeldin A, several Golgi proteins were detected in clusters that colocalize with the microtubule-organizing center in patient cells. All of these abnormalities were normalized in COG7-corrected patient fibroblasts. These results serve to better define the role of the COG complex in facilitating protein trafficking between the Golgi and ER and provide a diagnostic framework for the identification of CDG defects involving trafficking proteins.
authors.affiliation: Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA. rsteet@im.wustl.edu
authors.forename: Richard
authors.initials: R
authors.lastname: Steet
authors.forename: Stuart
authors.initials: S
authors.lastname: Kornfeld
mesh_terms: D033942:Adaptor Proteins, Vesicular Transport / Q000235:genetics / Q000378:metabolism*; D020126:Brefeldin A / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005033:Ethylmaleimide / Q000378:metabolism; D005347:Fibroblasts / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D037241:Mannose-Binding Lectins / Q000032:analysis / Q000378:metabolism*; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D008659:Metabolic Diseases / Q000235:genetics* / Q000378:metabolism; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism; D021381:Protein Transport; D050766:Qb-SNARE Proteins / Q000032:analysis / Q000378:metabolism*; D050767:Qc-SNARE Proteins / Q000032:analysis / Q000378:metabolism*
chemical_list: D033942:Adaptor Proteins, Vesicular Transport; C107588:BET1L protein, human; C512294:COG7 protein, human; C490972:GOSR1 protein, human; C084004:LMAN1 protein, human; D037241:Mannose-Binding Lectins; D008565:Membrane Proteins; D050766:Qb-SNARE Proteins; D050767:Qc-SNARE Proteins; D020126:Brefeldin A; D005033:Ethylmaleimide