AUTHOR=Giannakopoulou Natalia , Mendis Nilmini , Zhu Lei , Gruenheid Samantha , Faucher Sebastien P. , Le Moual Hervé 

TITLE=The Virulence Effect of CpxRA in Citrobacter rodentium Is Independent of the Auxiliary Proteins NlpE and CpxP

JOURNAL=Frontiers in Cellular and Infection Microbiology

VOLUME=8

YEAR=2018

URL=https://www.frontiersin.org/journals/cellular-and-infection-microbiology/articles/10.3389/fcimb.2018.00320

DOI=10.3389/fcimb.2018.00320

ISSN=2235-2988

ABSTRACT=<p><italic>Citrobacter rodentium</italic> is a murine pathogen used to model the intestinal infection caused by Enteropathogenic and Enterohemorrhagic <italic>Escherichia coli</italic> (EPEC and EHEC), two diarrheal pathogens responsible for morbidity and mortality in developing and developed countries, respectively. During infection, these bacteria must sense and adapt to the gut environment of the host. In order to adapt to changing environmental cues and modulate expression of specific genes, bacteria can use two-component signal transduction systems (TCS). We have shown that the deletion of the Cpx TCS in <italic>C. rodentium</italic> leads to a marked attenuation in virulence in C3H/HeJ mice. In <italic>E. coli</italic>, the Cpx TCS is reportedly activated in response to signals from the outer-membrane lipoprotein NlpE. We therefore investigated the role of NlpE in <italic>C. rodentium</italic> virulence. We also assessed the role of the reported negative regulator of CpxRA, CpxP. We found that as opposed to the Δ<italic>cpxRA</italic> strain, neither the Δ<italic>nlpE</italic>, Δ<italic>cpxP</italic> nor the Δ<italic>nlpE</italic>Δ<italic>cpxP</italic> strains were significantly attenuated, and had similar <italic>in vivo</italic> localization to wild-type <italic>C. rodentium</italic>. The <italic>in vitro</italic> adherence of the Cpx auxiliary protein mutants, Δ<italic>nlpE</italic>, Δ<italic>cpxP</italic>, Δ<italic>nlpE</italic>Δ<italic>cpxP</italic>, was comparable to wild-type <italic>C. rodentium</italic>, whereas the Δ<italic>cpxRA</italic> strain showed significantly decreased adherence. To further elucidate the mechanisms behind the contrasting virulence phenotypes, we performed microarrays in order to define the regulon of the Cpx TCS. We detected 393 genes differentially regulated in the Δ<italic>cpxRA</italic> strain. The gene expression profile of the Δ<italic>nlpE</italic> strain is strikingly different than the profile of Δ<italic>cpxRA</italic> with regards to the genes activated by CpxRA. Further, there is no clear inverse correlation in the expression pattern of the Δ<italic>cpxP</italic> strain in comparison to Δ<italic>cpxRA</italic>. Taken together, these data suggest that in these conditions, CpxRA activates gene expression in a largely NlpE- and CpxP-independent manner. Compared to wildtype, 161 genes were downregulated in the Δ<italic>cpxRA</italic> strain, while being upregulated or unchanged in the Cpx auxiliary protein deletion strains. This group of genes, which we hypothesize may contribute to the loss of virulence of Δ<italic>cpxRA</italic>, includes T6SS components, <italic>ompF</italic>, the regulator for colanic acid synthesis, and several genes involved in maltose metabolism.</p>