The mechanism of the phage-encoded protein antibiotic from ΦX174
Abstract
The historically important phage ΦX174 kills its host bacteria by encoding a 91-residue protein antibiotic called protein E. Using single-particle electron cryo–microscopy, we demonstrate that protein E bridges two bacterial proteins to form the transmembrane YES complex [MraY, protein E, sensitivity to lysis D (SlyD)]. Protein E inhibits peptidoglycan biosynthesis by obstructing the MraY active site leading to loss of lipid I production. We experimentally validate this result for two different viral species, providing a clear model for bacterial lysis and unifying previous experimental data. Additionally, we characterize the Escherichia coli MraY structure—revealing features of this essential enzyme—and the structure of the chaperone SlyD bound to a protein. Our structures provide insights into the mechanism of phage-mediated lysis and for structure-based design of phage therapeutics.
Additional Information
© 2023 the authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original US government works. We are particularly grateful to R. Young and T. Bernhardt for providing inspiration and feedback during the course of this project. We thank them, M. Kurosu, and D. Rees for critical feedback on the manuscript. We further thank R. Young for providing the ΔslyD strain. (Cryo)Electron microscopy was done in the Beckman Institute Resource Center for Transmission Electron Microscopy at Caltech. We are grateful to S. Chen for help with data collection and processing. This work was funded by the following: National Institutes of Health grant R01GM114611 (to W.M.C. and M.K.); National Institutes of Health grant DP1GM105385 (to W.M.C.); and the G. Harold and Leila Y. Mathers Foundation (to W.M.C.). Author contributions: Conceptualization: S.T. and W.M.C. Methodology: A.K.O., N.R., Y.E.L., S.T., H.G.Y., L.K., and W.M.C. Investigation: A.K.O., N.R., Y.E.L., S.T., H.G.Y., L.K., W.M.C. Visualization: A.K.O., N.R., S.T., and W.M.C. Funding acquisition: W.M.C. Project administration: W.M.C. Supervision: W.M.C. Writing – original draft: A.K.O., N.R., and W.M.C. Writing – review and editing: A.K.O., N.R., Y.E.L., S.T., H.G.Y., L.K., and W.M.C. Competing interests: Authors declare that they have no competing interests. Data and materials availability: All experimental data are available in the main text or the supplementary materials. Coordinates with experimental maps have been deposited to the RCSB or the EMDB for both the YESID21 (PDB ID 8G01, EMDB-29641) and YESΦX174 (PDB ID 8G02 and EMDB-29642) complexes.Attached Files
Supplemental Material - science.adg9091_movie_s1.zip
Supplemental Material - science.adg9091_sm.pdf
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Additional details
- Eprint ID
- 122507
- Resolver ID
- CaltechAUTHORS:20230725-857638000.85
- R01GM114611
- NIH
- DP1GM105385
- NIH
- G. Harold and Leila Y. Mathers Charitable Foundation
- Created
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2023-08-15Created from EPrint's datestamp field
- Updated
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2023-08-15Created from EPrint's last_modified field