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Journal of Virology, December 2002, p. 13083-13087, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.13083-13087.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Requirement for Vacuolar H+-ATPase Activity and Ca2+ Gradient during Entry of Rotavirus into MA104 Cells

Maria Elena Chemello, Olga Carolina Aristimuño, Fabián Michelangeli, and Marie-Christine Ruiz*

Laboratorio de Fisiología Gastrointestinal, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas, Venezuela

Received 29 April 2002/ Accepted 18 September 2002

The mechanism by which rotavirus and other nonenveloped viruses enter the cell is still not clear. We have proposed an endocytosis model where the critical step for virus uncoating and membrane permeabilization is the decrease in Ca2+ concentration in the endosome. In this paper, we monitored rotavirus entry by measuring {alpha}-sarcin-rotavirus coentry and infectivity in MA104 cells. The participation of endocytosis, acidification, and endosomal Ca2+ concentration on virus entry was studied by inhibiting the endosomal H+-ATPase with bafilomycin A1 and/or increasing the extracellular calcium reservoir by addition of 10 mM CaEGTA. Rotavirus-{alpha}-sarcin coentry was inhibited by bafilomycin A1 and by addition of 10 mM CaEGTA. These effects were additive. These substances induced a significant inhibition of infectivity without affecting virus binding and postentry steps. These results are compatible with the interpretation that bafilomycin A1 and CaEGTA block rotavirus penetration from the endosome into the cytoplasm and support our hypothesis of a Ca2+-dependent endocytosis model.

* Corresponding author. Mailing address: CBB-IVIC, Apartado 21827, 1020A Caracas, Venezuela. Phone: (58 212) 504 1164. Fax: (58 212) 504 1093. E-mail: mclr{at}cbb.ivic.ve.

Journal of Virology, December 2002, p. 13083-13087, Vol. 76, No. 24
0022-538X/02/$04.00+0     DOI: 10.1128/JVI.76.24.13083-13087.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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