Transposon-Directed Insertion-Site Sequencing Reveals Glycolysis Gene gpmA as Part of the H2O2 Defense Mechanisms in Escherichia coli

Myriam Roth; Emily C A Goodall; Karthik Pullela; Vincent Jaquet; Patrice François; Ian R Henderson; Karl-Heinz Krause

doi:10.3390/antiox11102053

Transposon-Directed Insertion-Site Sequencing Reveals Glycolysis Gene gpmA as Part of the H₂O₂ Defense Mechanisms in Escherichia coli

Antioxidants (Basel). 2022 Oct 18;11(10):2053. doi: 10.3390/antiox11102053.

Authors

Myriam Roth¹, Emily C A Goodall², Karthik Pullela², Vincent Jaquet^{1

3}, Patrice François⁴, Ian R Henderson², Karl-Heinz Krause¹

Affiliations

¹ Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, 1211 Geneva, Switzerland.
² Institute for Molecular Bioscience, University of Queensland, St. Lucia, Brisbane, QLD 4072, Australia.
³ READS Unit, Faculty of Medicine, University of Geneva, 1211 Geneva, Switzerland.
⁴ Genomic Research Laboratory, Infectious Diseases Service, University Hospitals of Geneva, University Medical Center, Michel-Servet 1, 1211 Geneva, Switzerland.

Abstract

Hydrogen peroxide (H₂O₂) is a common effector of defense mechanisms against pathogenic infections. However, bacterial factors involved in H₂O₂ tolerance remain unclear. Here we used transposon-directed insertion-site sequencing (TraDIS), a technique allowing the screening of the whole genome, to identify genes implicated in H₂O₂ tolerance in Escherichia coli. Our TraDIS analysis identified 10 mutants with fitness defect upon H₂O₂ exposure, among which previously H₂O₂-associated genes (oxyR, dps, dksA, rpoS, hfq and polA) and other genes with no known association with H₂O₂ tolerance in E. coli (corA, rbsR, nhaA and gpmA). This is the first description of the impact of gpmA, a gene involved in glycolysis, on the susceptibility of E. coli to H₂O₂. Indeed, confirmatory experiments showed that the deletion of gpmA led to a specific hypersensitivity to H₂O₂ comparable to the deletion of the major H₂O₂ scavenger gene katG. This hypersensitivity was not due to an alteration of catalase function and was independent of the carbon source or the presence of oxygen. Transcription of gpmA was upregulated under H₂O₂ exposure, highlighting its role under oxidative stress. In summary, our TraDIS approach identified gpmA as a member of the oxidative stress defense mechanism in E. coli.

Keywords: E. coli; H2O2; Tn-seq; TraDIS; gpmA; phophoglycerate mutase.

Grants and funding

This study was granted by Swiss National Science Foundation to Karl-Heinz Krause (Funding number 31003A-179478). The TraDIS work was supported by University of Queensland funding to Ian Henderson.