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Journal of Virology, April 2005, p. 5017-5026, Vol. 79, No. 8
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.8.5017-5026.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Penetration of Enveloped Double-Stranded RNA Bacteriophages 13 and 6 into Pseudomonas syringae Cells

Rimantas Daugelaviius,^1,2 Virginija Cvirkait,^1,2 Aura Gaidelyt,^1,2 Elena Bakien,² Rasa Gabrnait-Verkhovskaya,^1, and Dennis H. Bamford^1*

Department of Biological and Environmental Sciences and Institute of Biotechnology, University of Helsinki, Helsinki, Finland,¹ Department of Biochemistry and Biophysics, Vilnius University, Vilnius, Lithuania²

Received 20 September 2004/ Accepted 22 November 2004

Bacteriophages 6 and 13 are related enveloped double-stranded RNA viruses that infect gram-negative Pseudomonas syringae cells. 6 uses a pilus as a receptor, and 13 attaches to the host lipopolysaccharide. We compared the entry-related events of these two viruses, including receptor binding, envelope fusion, peptidoglycan penetration, and passage through the plasma membrane. The infection-related events are dependent on the multiplicity of infection in the case of 13 but not with 6. A temporal increase of host outer membrane permeability to lipophilic ions was observed from 1.5 to 4 min postinfection in both virus infections. This enhanced permeability period coincided with the fast dilution of octadecyl rhodamine B-labeled virus-associated lipid molecules. This result is in agreement with membrane fusion, and the presence of temporal virus-derived membrane patches on the outer membrane. Similar to 6, 13 contains a thermosensitive lytic enzyme involved in peptidoglycan penetration. The phage entry also caused a limited depolarization of the plasma membrane. Inhibition of host respiration considerably decreased the efficiency of irreversible virus binding and membrane fusion. An active role of cell energy metabolism in restoring the infection-induced defects in the cell envelope was also observed.

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* Corresponding author. Mailing address: Viikki Biocenter, P.O. Box 56 (Viikinkaari 5) FI-00014, University of Helsinki, Helsinki, Finland. Phone: 358-9-191-59100. Fax: 358-9-191-59098. E-mail: dennis.bamford{at}helsinki.fi.

Present address: Department of Applied Biology, University of Helsinki, Helsinki, Finland.

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Journal of Virology, April 2005, p. 5017-5026, Vol. 79, No. 8
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.8.5017-5026.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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