Dynamin is involved in human epithelial cell vacuolation caused by the Helicobacter pylori–produced cytotoxin VacA

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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.

Authors

Junko Suzuki, Hirohide Ohnsihi, Hiroshi Shibata, Akihiro Wada, Toshiya Hirayama, Taroh Iiri, Namiki Ueda, Chiho Kanamaru, Tomohiro Tsuchida, Hirosato Mashima, Hiroshi Yasuda, Toshiro Fujita