Structural Modeling of Cell Wall Peptidase CwpFM (EntFM) Reveals Distinct Intrinsically Disordered Extensions Specific to Pathogenic Bacillus cereus Strains

Abstract

The emergence of B. cereus as an opportunistic food-borne pathogen has intensified the need to distinguish strains of public health concern. The heterogeneity of the diseases associated with B. cereus infections emphasizes the versatility of these bacteria strains to colonize their host. Nevertheless, the molecular basis of these differences remains unclear. Several toxins are involved in virulence, particularly in gastrointestinal disorders, but there are currently no biological markers able to differentiate pathogenic from harmless strains. We have previously shown that CwpFM is a cell wall peptidase involved in B. cereus virulence. Here, we report a sequence/structure/function characterization of 39 CwpFM sequences, chosen from a collection of B. cereus with diverse virulence phenotypes, from harmless to highly pathogenic strains. CwpFM is homology-modeled in silico as an exported papain-like endopeptidase, with an N-terminal end composed of three successive bacterial Src Homology 3 domains (SH3b1–3) likely to control protein–protein interactions in signaling pathways, and a C-terminal end that contains a catalytic NLPC_P60 domain primed to form a competent active site. We confirmed in vitro that CwpFM is an endopeptidase with a moderate peptidoglycan hydrolase activity. Remarkably, CwpFMs from pathogenic strains harbor a specific stretch of twenty residues intrinsically disordered, inserted between the SH3b3 and the catalytic NLPC_P60 domain. This strongly suggests this linker as a marker of differentiation between B. cereus strains. We believe that our findings improve our understanding of the pathogenicity of B. cereus while advancing both clinical diagnosis and food safety.

Bacillus cereus Induces Severe Infections in Preterm Neonates: Implication at the Hospital and Human Milk Bank Level

“…The Mfd protein repairs damage to bacterial DNA caused by nitrogen stress during the host’s immune response to infection [ 54 , 55 ]. CwpFM is involved in the adhesion to epithelial cells and biofilm formation, and CwpFM from pathogenic strains possess structural specificities [ 56 , 57 ].…”

New genetic biomarkers to differentiate non-pathogenic from clinically relevant Bacillus cereus strains

“…Moreover, these toxins do not appear to be suitable markers for strains causing non-gastrointestinal infections [9]. B. cereus produces other toxins such as haemolysin II (HlyII), the metalloproteases InhA1, InhA2 and the cell wall peptidase FM (CwpFM), which may also be involved in pathogenicity [14][15][16][17][18]. The emetic form of B. cereus food poisoning is caused by the peptide cereulide [19], which represent less than 1% of the FBO strains of B. cereus [1,19,20].…”

The pathogenic biomarker alcohol dehydrogenase protein is involved in Bacillus cereus virulence and survival against host innate defence

“…In addition, other factors such as HlyII, InhA1, CwpFM or Mfd have been implicated in B . cereus resistance against the host immune system [ 15 – 21 ]. These toxins provide an indication of the strain toxicity potential [ 13 , 22 – 24 ].…”