Abstract

Enteropathogenic Escherichia coli (EPEC) is a diarrheagenic pathogen that afflicts infants in developing countries. The most important virulence trait of EPEC is its ability to intimately adhere to cells in the small intestine, and to elicit diarrhea. The alternative sigma factor RpoS is involved in the virulence of several bacterial species. RpoS coordinates the general stress response and accumulates in cells under stress or in the stationary phase. RpoS levels differ across E. coli strains. High-RpoS strains are highly resistant to environmental stresses, but usually display low nutritional competence, while low-RpoS strains show the opposite phenotype. Here we investigated whether RpoS plays a role in the virulence and fitness of two different EPEC strains, E2348/69 and LRT9. A rpoS null mutation had a small positive effect on LRT9 adherence to epithelial cells, but the expression of the EPEC adhesins BfpA and intimin was not significantly affected by the mutation. E2348/69 adherence was not significantly affected by the rpoS mutation. The intrinsic level of RpoS was higher in LRT9 than in E2348/69 while the latter adhered more strongly and expressed higher levels of the adhesin BfpA than the former. Knockout of rpoS strongly impaired resistance to oxidative, osmotic and acid stress in both E2348/69 and LRT9. However, strain E2348/69 was significantly more sensitive to oxidative stress than LRT9. Finally, competition assays showed that the rpoS mutant of LRT9 displayed higher fitness under continuous culture than its isogenic wild-type strain, while E2348/69 outcompeted its rpoS mutant. In conclusion, RpoS plays mostly a positive role in EPEC biology and at least in the case of strain E2348/69 it is not constrained by the trade-off between vegetative growth and stress resistance.

Highlights

  • The sigma factor RpoS is the master regulator of the general stress response in E. coli [1]

  • RpoS role in enteropathogenic Escherichia coli and elsewhere to study the role of regulatory genes in Enteropathogenic Escherichia coli (EPEC) adherence [28, 30, 36]. Both E2348/69 and LRT9 rpoS::Tn10 mutants displayed slow bubbling in the presence of hydrogen peroxide and were considerably more sensitive to oxidative, osmotic and acid stress (Fig 1), as expected for rpoS-negative strains

  • The Str resistance is due to the presence of the plasmid pE2348-2 which carries the strAB genes [37], while the Nal resistance was deliberately selected in the original E2348/69 StrR strain [38]

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Summary

Introduction

The sigma factor RpoS is the master regulator of the general stress response in E. coli [1]. HosA encodes a transcriptional regulator that belongs to the SlyA family; pad codes for a phenylacrylic acid decarboxylase, that confers resistance to phenylacrylic acids and yclC encodes the C subunit of a phenolic acid decarboxylase. Complementation with pNP5 did not significantly alter the behavior of EPEC regarding those phenotypes These results indicate that the 2.9 Kb region downstream of rpoS does not exert any influence on EPEC adherence. These findings are in agreement with [62] that showed that hosA, which is located immediately downstream of rpoS in EPEC and EHEC, did not affect type III secretion, LEE1 and LEE4 regulation, or the ability of E2348/69 to form attaching-and-effacing lesions on intestinal epithelial cells

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