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Published June 14, 2011 | Supplemental Material + Published
Journal Article Open

Alternative mechanism for bacteriophage adsorption to the motile bacterium Caulobacter crescentus

Abstract

2D and 3D cryo-electron microscopy, together with adsorption kinetics assays of ϕCb13 and ϕCbK phage-infected Caulobacter crescentus, provides insight into the mechanisms of infection. ϕCb13 and ϕCbK actively interact with the flagellum and subsequently attach to receptors on the cell pole. We present evidence that the first interaction of the phage with the bacterial flagellum takes place through a filament on the phage head. This contact with the flagellum facilitates concentration of phage particles around the receptor (i.e., the pilus portals) on the bacterial cell surface, thereby increasing the likelihood of infection. Phage head filaments have not been well characterized and their function is described here. Phage head filaments may systematically underlie the initial interactions of phages with their hosts in other systems and possibly represent a widespread mechanism of efficient phage propagation.

Additional Information

© 2011 National Academy of Sciences. Freely available online through the PNAS open access option. Edited by Sankar Adhya, National Institutes of Health, National Cancer Institute, Bethesda, MD, and approved April 22, 2011 (received for review August 20, 2010). Published online before print May 25, 2011. We thank Dr. Jens M. Holl and Mr. Grant M. Williams for valuable help with manuscript preparation, graphic arts, and helpful discussions; Dr. Ryland Young and Dr. Ian Molineux for advice and mentoring on phage biology; Ms. Jeannette Taylor, Ms. Hong Yi, Dr. Wei Zhang, Dr. Ozan Ugurlu, and Mr. Chris Frethem for assistance during cryo-EM data collection; and Ms. Sabine Pruggnaller for providing the Amira toolbox for 3D segmentation. This work was supported in part by Emory University, Children's Healthcare of Atlanta, and the Georgia Research Alliance (to E.R.W.); and Human Frontier Science Program Grant RGP0051 (to P.H.V. and E.R.W.). The work at California Institute of Technology was supported by National Institutes of Health Grant P01 GM066521 (to G.J.J.) and a gift to Caltech from the Gordon and Betty Moore Foundation. Parts of this work were carried out in the Institute of Technology Characterization Facility, University of Minnesota, which receives partial support from National Science Foundation through the Materials Research Science & Engineering Centers (MRSEC) program. Author contributions: R.C.G.-F. and E.R.W. designed research; R.C.G.-F., M.G., and E.R.W. performed research; P.H.V., B.E., J.S.P., and G.J.J. contributed new reagents/analytic tools; R.C.G.-F. and E.R.W. analyzed data; R.C.G.-F. and E.R.W. wrote the paper; and P.H.V., B.E., and J.S.P. contributed bacterial strains and phage stocks.

Attached Files

Published - GuerreroFerreira2011p14282P_Natl_Acad_Sci_Usa.pdf

Supplemental Material - pnas.201012388SI.pdf

Supplemental Material - sm01.mp4

Supplemental Material - sm02.mp4

Supplemental Material - sm03.mp4

Supplemental Material - sm04.mp4

Supplemental Material - sm05.mp4

Supplemental Material - sm06.mp4

Supplemental Material - sm07.mp4

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