title: Bacterial ADP-ribosyltransferase with a substrate specificity of the rho protein disassembles the Golgi apparatus in Vero cells and mimics the action of brefeldin A.
abstract: Epidermal-cell differentiation inhibitor (EDIN) is an exoenzyme produced by Staphylococcus aureus that catalyzes the ADP-ribosylation of rho proteins, members of the small GTP-binding protein family. In this study we demonstrate that EDIN induces a rapid morphological change in the Golgi structure of monkey kidney Vero cells that is similar to the changes elicited by brefeldin A (BFA). Treatment of Vero cells with EDIN resulted in a rapid disappearance of N-7-(4-nitrobenzo-2-oxa-1,3-diazole)-6-aminocaproylsphingosine, a 110-kDa protein (beta-COP, coat protein), and mannosidase II from the Golgi structure. Lower doses of EDIN and BFA had a synergistic effect on the redistribution of the Golgi markers. The similarities in the effects of EDIN and BFA in Vero cells also include the EDIN- or BFA-mediated protection of Vero cells from ricin cytotoxicity and prevention of the effects of EDIN or BFA on the distribution of Golgi markers by the pretreatment of Vero cells with guanosine 5'-[gamma-thio]triphosphate or forskolin. Incubation of a Vero-cell homogenate with [32P]NAD+ and EDIN in vitro resulted in the appearance of a labeled band with an apparent molecular mass of 22 kDa. The morphological change of the Golgi structure induced by EDIN was inhibited by nicotinamide, an inhibitor of EDIN-catalyzed ADP-ribosylation. Thus these data suggest that a rho protein is involved in the membrane trafficking between the Golgi and the endoplasmic reticulum of Vero cells and that this rho protein may be a target shared by EDIN and BFA.
authors.affiliation: Department of Microbiology, Uniformed Services University of the Health Sciences, Bethesda, MD 20814-4799.
authors.forename: M
authors.initials: M
authors.lastname: Sugai
authors.forename: C H
authors.initials: CH
authors.lastname: Chen
authors.forename: H C
authors.initials: HC
authors.lastname: Wu
mesh_terms: D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives / Q000378:metabolism; D000246:Adenosine Diphosphate Ribose / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology; D015415:Biomarkers; D020126:Brefeldin A; D002518:Ceramides / Q000378:metabolism; D020756:Coatomer Protein; D005576:Colforsin / Q000494:pharmacology; D003517:Cyclopentanes / Q000494:pharmacology*; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000494:pharmacology; D008361:Mannosidases / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D012234:Rho Factor / Q000378:metabolism*; D013211:Staphylococcus aureus; D013379:Substrate Specificity; D014709:Vero Cells
chemical_list: D001426:Bacterial Proteins; D015415:Biomarkers; D002518:Ceramides; D020756:Coatomer Protein; D003517:Cyclopentanes; D008869:Microtubule-Associated Proteins; D012234:Rho Factor; C067120:epidermal cell differentiation inhibitor, Staphylococcus aureus; D005576:Colforsin; D020126:Brefeldin A; D000246:Adenosine Diphosphate Ribose; D016244:Guanosine 5'-O-(3-Thiotriphosphate); C037974:N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl sphingosine; D011065:Poly(ADP-ribose) Polymerases; D008361:Mannosidases; C038829:mannosyl-oligosaccharide 1,3 - 1,6-alpha-mannosidase; D009327:4-Chloro-7-nitrobenzofurazan