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

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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, 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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