Various forms of cancer have been linked to the carcinogenic activities of microorganisms1–3. The virulent gene island polyketide synthase (pks) produces the secondary metabolite colibactin, a genotoxic molecule(s) causing double-stranded DNA breaks4 and enhanced colorectal cancer development5,6. Colibactin biosynthesis involves a prodrug resistance strategy where an N-terminal prodrug scaffold (precolibactin) is assembled, transported into the periplasm and cleaved to release the mature product7–10. Here, we show that ClbM, a multidrug and toxic compound extrusion (MATE) transporter, is a key component involved in colibactin activity and transport. Disruption of clbM attenuated pks+ E. coli-induced DNA damage in vitro and significantly decreased the DNA damage response in gnotobiotic Il10−/− mice. Colonization experiments performed in mice or zebrafish animal models indicate that clbM is not implicated in E. coli niche establishment. The X-ray structure of ClbM shows a structural motif common to the recently described MATE family. The 12-transmembrane ClbM is characterized as a cation-coupled antiporter, and residues important to the cation-binding site are identified. Our data identify ClbM as a precolibactin transporter and provide the first structure of a MATE transporter with a defined and specific biological function.
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This work was supported by funding from R01 DK73338 (C.J.), R01DK47700 (C.J.), R01 GM0865700 (S.D.B.) and by the University of Florida (S.D.B.) and the UF DoM Gatorade Fund (C.J.). The authors thank V. de Crecy-Lagard for discussions.
The authors declare no competing financial interests.
Supplementary Figures 1–9 and Tables 1,2. (PDF 1329 kb)
NC101-colonized, fixed 6-d.p.f. zebrafish. Reconstruction from z-stack images; DAPI stained whole mount zebrafish with tdTomato-labelled bacteria. (MOV 1444 kb)
ΔclbM-colonized, fixed 6-d.p.f. zebrafish. Reconstruction from z-stack images; DAPI stained whole mount zebrafish with tdTomato-labelled bacteria. (MOV 1559 kb)
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Mousa, J., Yang, Y., Tomkovich, S. et al. MATE transport of the E. coli-derived genotoxin colibactin. Nat Microbiol 1, 15009 (2016). https://doi.org/10.1038/nmicrobiol.2015.9
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