RRC ID 41335
Author Suzuki J, Ohnsihi H, Shibata H, Wada A, Hirayama T, Iiri T, Ueda N, Kanamaru C, Tsuchida T, Mashima H, Yasuda H, Fujita T.
Title Dynamin is involved in human epithelial cell vacuolation caused by the Helicobacter pylori-produced cytotoxin VacA.
Journal J. Clin. Invest.
Abstract The Helicobacter pylori-produced cytotoxin VacA induces intracellular vacuolation. To elucidate the molecular mechanism of vacuole formation by VacA, we examined the participation of dynamin, a GTPase functioning in intracellular vesicle formation, in human HeLa cells. Immunocytochemistry revealed that endogenous dynamin was localized to vacuoles induced by VacA. In cells transiently transfected with a GTPase-defective (dominant-negative) dynamin mutant, VacA failed to induce vacuolation. In contrast, VacA did induce vacuolation in cells transiently transfected with wild-type dynamin. Furthermore, under VacA treatment, neutral red dye uptake, a parameter of VacA-induced vacuolation, was inhibited in cells stably transfected with the dominant-negative dynamin mutant. In contrast, uptake was markedly enhanced in cells stably transfected with wild-type dynamin. Moreover, VacA cytopathic effects on the viability of HeLa cells were inhibited in cells stably transfected with dominant-negative dynamin-1. Sequential immunocytochemical observation confirmed that expression of dominant-negative dynamin did not affect VacA attachment to or internalization into HeLa cells. We suggest that dynamin is involved in the intracellular vacuolation induced by VacA.
Volume 107(3)
Pages 363-70
Published 2001-2
DOI 10.1172/JCI10254
PMID 11160160
PMC PMC199191
MeSH Bacterial Proteins / antagonists & inhibitors Bacterial Proteins / biosynthesis* Cell Survival Cytotoxins / biosynthesis* Duodenal Diseases / microbiology Dynamin I Dynamins Epithelial Cells / metabolism* Fluorescent Antibody Technique GTP Phosphohydrolases / genetics GTP Phosphohydrolases / metabolism* HeLa Cells Helicobacter pylori / metabolism* Humans Plasmids Transfection
IF 13.251
Times Cited 33
WOS Category MEDICINE, RESEARCH & EXPERIMENTAL
Resource
Human and Animal Cells