title: A non-invasive method for studying viral DNA delivery to bacteria reveals key requirements for phage SPP1 DNA entry in Bacillus subtilis cells.
abstract: Bacteriophages use most frequently a tail apparatus to create a channel across the entire bacterial cell envelope to transfer the viral genome to the host cell cytoplasm, initiating infection. Characterization of this critical step remains a major challenge due to the difficulty to monitor DNA entry in the bacterium and its requirements. In this work we developed a new method to study phage DNA entry that has the potential to be extended to many tailed phages. Its application to study genome delivery of bacteriophage SPP1 into Bacillus subtilis disclosed a key role of the host cell membrane potential in the DNA entry process. An energized B. subtilis membrane and a millimolar concentration of calcium ions are shown to be major requirements for SPP1 DNA entry following the irreversible binding of phage particles to the receptor YueB.
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Sofia
authors.initials: S
authors.lastname: Fernandes
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Saclay, UMR 9198, 91198 Gif-sur-Yvette cedex, France; Unit of Molecular and Structural Virology (VMS), UPR3296 CNRS, Campus CNRS, 91198 Gif-sur-Yvette cedex, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Labarde
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal.
authors.forename: Catarina
authors.initials: C
authors.lastname: Baptista
authors.affiliation: Unit of Molecular and Structural Virology (VMS), UPR3296 CNRS, Campus CNRS, 91198 Gif-sur-Yvette cedex, France.
authors.forename: Lina
authors.initials: L
authors.lastname: Jakutytè
authors.affiliation: Institute for Integrative Biology of the Cell (I2BC), CEA, CNRS, Université Paris-Saclay, UMR 9198, 91198 Gif-sur-Yvette cedex, France; Unit of Molecular and Structural Virology (VMS), UPR3296 CNRS, Campus CNRS, 91198 Gif-sur-Yvette cedex, France.
authors.forename: Paulo
authors.initials: P
authors.lastname: Tavares
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculty of Pharmacy, Universidade de Lisboa, Av. Prof. Gama Pinto, 1649-003 Lisboa, Portugal. Electronic address: csaojose@ff.ul.pt.
authors.forename: Carlos
authors.initials: C
authors.lastname: São-José
mesh_terms: D001412:Bacillus subtilis / Q000378:metabolism / Q000821:virology*; D001435:Bacteriophages / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D004279:DNA, Viral; D018014:Gene Transfer Techniques; D016679:Genome, Viral; D006096:Gramicidin / Q000494:pharmacology; D008564:Membrane Potentials; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging; D014161:Transduction, Genetic; D053586:Virus Internalization / Q000187:drug effects; D057646:Virus Uncoating
chemical_list: D004279:DNA, Viral; D006096:Gramicidin; D002118:Calcium
keywords: B. subtilis; Bacteriophage; Calcium; Membrane potential; SPP1; Virus entry; YueB