Colicin-mediated transport of DNA through the iron transporter FepA

Cohen-Khait, R.; Harmalkar, A.; Pham, P.; Webby, M. N.; Housden, N. G.; Elliston, E.; Hopper, J. T.; Mohammed, S.; Robinson, C. V.; Gray, J. J.; Kleanthous, C.

doi:10.1101/2021.05.11.443673

bioRxiv · 10.1101/2021.05.11.443673

Colicin-mediated transport of DNA through the iron transporter FepA

Cohen-Khait, R.·Harmalkar, A.·Pham, P.·Webby, M. N.·Housden, N. G.·Elliston, E.·Hopper, J. T.·Mohammed, S.·Robinson, C. V.·Gray, J. J.·Kleanthous, C.

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Abstract

Colicins are protein antibiotics used by bacteria to eliminate competing Escherichia coli. Colicins frequently exploit outer membrane (OM) nutrient transporters to penetrate through the strictly impermeable bacterial cellular envelope. Here, applying live-cell fluorescence imaging we were able to follow colicin B (ColB) into E. coli and localize it within the periplasm. We further demonstrate that single-stranded DNA coupled to ColB is also transported into the periplasm, emphasizing that the import routes of colicins can be exploited to carry large cargo molecules into bacteria. Moreover, we characterize the molecular mechanism of ColB association with its OM receptor FepA, applying a combination of photo-activated crosslinking, mass spectrometry, and structural modeling. We demonstrate that complex formation is coincident with a large-scale conformational change in the colicin. Finally In vivo crosslinking experiments and supplementary simulations of the translocation process indicate that part of the colicin engages active transport by disguising itself to part of the cellular receptor.

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Cohen-Khait, R., Harmalkar, A., Pham, P., Webby, M. N., Housden, N. G., Elliston, E., Hopper, J. T., Mohammed, S., Robinson, C. V., Gray, J. J., Kleanthous, C.. 2021-05-11. Colicin-mediated transport of DNA through the iron transporter FepA. https://doi.org/10.1101/2021.05.11.443673

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