Model Colibactins Exhibit Human Cell Genotoxicity in the Absence of Host Bacteria

Emilee E. Shine, Mengzhao Xue, Jaymin R. Patel, Alan Healy, Yulia V. Surovtseva, Seth B. Herzon, Jason M. Crawford

    Research output: Contribution to journalArticle

    Abstract

    Colibactins are genotoxic secondary metabolites produced in select Enterobacteriaceae, which induce downstream DNA double-strand breaks (DSBs) in human cell lines and are thought to promote the formation of colorectal tumors. Although key structural and functional features of colibactins have been elucidated, the full molecular mechanisms regulating these phenotypes remain unknown. Here, we demonstrate that free model colibactins induce DSBs in human cell cultures and do not require delivery by host bacteria. Through domain-targeted editing, we demonstrate that a subset of native colibactins generated from observed module skipping in the nonribosomal peptide synthetase-polyketide synthase (NRPS-PKS) biosynthetic assembly line share DNA alkylation phenotypes with the model colibactins in vitro. However, module skipping eliminates the strong DNA interstrand cross-links formed by the wild-type pathway in cell culture. This product diversification during the modular NRPS-PKS biosynthesis produces a family of metabolites with varying observed mechanisms of action (DNA alkylation versus cross-linking) in cell culture. The presence of membranes separating human cells from model colibactins attenuated genotoxicity, suggesting that membrane diffusion limits colibactin activity and could account for the reported bacterium-human cell-to-cell contact phenotype. Additionally, extracellular supplementation of the colibactin resistance protein ClbS was able to intercept colibactins in an Escherichia coli-human cell transient infection model. Our studies demonstrate that free model colibactins recapitulate cellular phenotypes associated with module-skipped products in the native colibactin pathway and define specific protein domains that are required for efficient DNA interstrand cross-linking in the native pathway.

    Original languageEnglish (US)
    Pages (from-to)3286-3293
    Number of pages8
    JournalACS Chemical Biology
    Volume13
    Issue number12
    DOIs
    StatePublished - Dec 21 2018

    Fingerprint

    Bacteria
    Cells
    Cell culture
    Peptide Synthases
    DNA
    Polyketide Synthases
    Phenotype
    Cell Culture Techniques
    Alkylation
    Metabolites
    colibactin
    Membranes
    Double-Stranded DNA Breaks
    Biosynthesis
    Enterobacteriaceae
    Escherichia coli
    Tumors
    Colorectal Neoplasms
    Proteins
    Cell Line

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Medicine

    Cite this

    Shine, E. E., Xue, M., Patel, J. R., Healy, A., Surovtseva, Y. V., Herzon, S. B., & Crawford, J. M. (2018). Model Colibactins Exhibit Human Cell Genotoxicity in the Absence of Host Bacteria. ACS Chemical Biology, 13(12), 3286-3293. https://doi.org/10.1021/acschembio.8b00714

    Model Colibactins Exhibit Human Cell Genotoxicity in the Absence of Host Bacteria. / Shine, Emilee E.; Xue, Mengzhao; Patel, Jaymin R.; Healy, Alan; Surovtseva, Yulia V.; Herzon, Seth B.; Crawford, Jason M.

    In: ACS Chemical Biology, Vol. 13, No. 12, 21.12.2018, p. 3286-3293.

    Research output: Contribution to journalArticle

    Shine, EE, Xue, M, Patel, JR, Healy, A, Surovtseva, YV, Herzon, SB & Crawford, JM 2018, 'Model Colibactins Exhibit Human Cell Genotoxicity in the Absence of Host Bacteria', ACS Chemical Biology, vol. 13, no. 12, pp. 3286-3293. https://doi.org/10.1021/acschembio.8b00714
    Shine EE, Xue M, Patel JR, Healy A, Surovtseva YV, Herzon SB et al. Model Colibactins Exhibit Human Cell Genotoxicity in the Absence of Host Bacteria. ACS Chemical Biology. 2018 Dec 21;13(12):3286-3293. https://doi.org/10.1021/acschembio.8b00714
    Shine, Emilee E. ; Xue, Mengzhao ; Patel, Jaymin R. ; Healy, Alan ; Surovtseva, Yulia V. ; Herzon, Seth B. ; Crawford, Jason M. / Model Colibactins Exhibit Human Cell Genotoxicity in the Absence of Host Bacteria. In: ACS Chemical Biology. 2018 ; Vol. 13, No. 12. pp. 3286-3293.
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