Cellular vacuoles induced by Helicobacter pylori originate from late endosomal compartments.
Pathogenic strains of Helicobacter pylori cause progressive vacuolation and death of epithelial cells. To identify the nature of vacuoles, the distribution of markers of various membrane traffic compartments was studied. Vacuoles derive from the endocytic pathway since they include the fluid-phase marker Lucifer yellow. Early endosome markers such as rab5, transferrin, and transferrin receptor, as well as the lysosomal hydrolase cathepsin D, are excluded from these structures. In contrast, the vacuolar membrane is specifically stained by affinity-purified antibodies against rab7, a small GTPase, localized to late endosomal compartments. The labeling of rab7 on vacuolar membranes increases as vacuolation progresses, without a concomitant increase of cellular rab7. Cell vacuolation is inhibited by the microtubule-depolymerizing agents nocodazole and colchicine. Taken together, these findings indicate that the vacuoles specifically originate from late endosomal compartments.
Papini
E
E
Dipartimento di Scienze Biomediche, Università di Padova, Italy.
de Bernard
M
M
Milia
E
E
Bugnoli
M
M
Zerial
M
M
Rappuoli
R
R
Montecucco
C
C
D000225:Adenine / Q000031:analogs & derivatives / Q000494:pharmacology; D015415:Biomarkers / Q000032:analysis; D002402:Cathepsin D / Q000032:analysis; D003078:Colchicine / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D019204:GTP-Binding Proteins / Q000032:analysis; D006367:HeLa Cells; D016480:Helicobacter pylori / Q000472:pathogenicity*; D006801:Humans; D015151:Immunoblotting; D007546:Isoquinolines; D007700:Kinetics; D015739:Nocodazole / Q000494:pharmacology; D011990:Receptors, Transferrin / Q000032:analysis; D014617:Vacuoles / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*
D015415:Biomarkers; D007546:Isoquinolines; D011990:Receptors, Transferrin; C025946:3-methyladenine; C017475:lucifer yellow; D002402:Cathepsin D; D019204:GTP-Binding Proteins; D000225:Adenine; D015739:Nocodazole; D003078:Colchicine