Junko Suzuki, Hirohide Ohnsihi, Hiroshi Shibata, Akihiro Wada, Toshiya Hirayama, Taroh Iiri, Namiki Ueda, Chiho Kanamaru, Tomohiro Tsuchida, Hirosato Mashima, Hiroshi Yasuda, Toshiro Fujita
J Clin Invest.
2001;
107(3):363–370
doi:10.1172/JCI10254
This article Copyright © 2001, The American Society for Clinical Investigation
Abstract
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T
he 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.
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