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bioRxiv · 10.1101/2024.01.30.576698

The Fanconi anemia pathway repairs colibactin-induced DNA interstrand cross-links

Altshuller, M.·He, X.·MacKrell, E. J.·Wernke, K. M.·Wang, J. W. H.·Selles-Baiget, S.·Wang, T.-Y.·Chou, T.-F.·Duxin, J. P.·Balskus, E. P.·Herzon, S. B.·Semlow, D.
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Abstract

Colibactin is a secondary metabolite produced by bacteria present in the human gut and is implicated in the progression of colorectal cancer and inflammatory bowel disease. This genotoxin alkylates deoxyadenosines on opposite strands of host cell DNA to produce DNA interstrand cross-links (ICLs) that block DNA replication. While cells have evolved multiple mechanisms to resolve ("unhook") ICLs encountered by the replication machinery, little is known about which of these pathways promote resistance to colibactin-induced ICLs. Here, we use Xenopus egg extracts to investigate replication-coupled repair of plasmids engineered to contain site-specific colibactin-ICLs. We show that replication fork stalling at a colibactin-ICL leads to replisome disassembly and activation of the Fanconi anemia ICL repair pathway, which unhooks the colibactin-ICL through nucleolytic incisions. These incisions generate a DNA double-strand break intermediate in one sister chromatid, which can be repaired by homologous recombination, and a monoadduct ("ICL remnant") in the other. Our data indicate that translesion synthesis past the colibactin-ICL remnant depends on Pol{eta} and a Pol{kappa}-REV1-Pol{zeta} polymerase complex. Although translesion synthesis past colibactin-induced DNA damage is frequently error-free, it can introduce T>N point mutations that partially recapitulate the mutation signature associated with colibactin exposure in vivo. Taken together, our work provides a biochemical framework for understanding how cells tolerate a naturally-occurring and clinically-relevant ICL.

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Altshuller, M., He, X., MacKrell, E. J., Wernke, K. M., Wang, J. W. H., Selles-Baiget, S., Wang, T.-Y., Chou, T.-F., Duxin, J. P., Balskus, E. P., Herzon, S. B., Semlow, D.. 2024-01-30. The Fanconi anemia pathway repairs colibactin-induced DNA interstrand cross-links. https://doi.org/10.1101/2024.01.30.576698

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