Analysis of the E Regulon in Crohn’s Disease-Associated Escherichia coli Revealed Involvement of the waaWVL Operon in Biofilm Formation

Ileal lesions of patients with Crohn’s disease are colonized by adherent-invasive Escherichia coli (AIEC), which is able to adhere to and to invade intestinal epithelial cells (IEC), to replicate within macrophages, and to form biofilms on the surface of the in-testinal mucosa. Previous analyses indicated the involvement of theE pathway in AIEC-IEC interaction, as well as in biofilm formation, withE pathway inhibition leading to an impaired ability of AIEC to colonize the intestinal mucosa and to form bio-films. The aim of this study was to characterize theE regulon of AIEC strain LF82 in order to identify members involved in AIEC phenotypes. Using comparative in silico analysis of theE regulon, we identified the waaWVL operon as a new member of theE regulon in reference AIEC strain LF82. We determined that the waaWVL operon is involved in AIEC lipopolysaccharide structure and composition, and the waaWVL operon was found to be essential for AIEC strains to produce biofilm and to colo-nize the intestinal mucosa. IMPORTANCE An increased prevalence of adherent-invasive Escherichia coli (AIEC) bacteria was previously observed in the intestinal mucosa of Crohn’s disease (CD) patients, and clinical observations revealed bacterial biofilms associated with the mucosa of CD pa-tients. Here, analysis of theE regulon in AIEC and commensal E. coli identified 12 genes controlled byE only in AIEC. Among them, WaaWVL factors were found to play an essential role in biofilm formation and mucosal colonization by AIEC. In additio