A trimeric autotransporter enhances biofilm cohesiveness in Yersinia pseudotuberculosis but not in Yersinia pestis

Abstract

Cohesion of biofilms made by Yersinia pestis and Yersinia pseudotuberculosis (Yptb) has been attributed solely to an extracellular polysaccharide matrix encoded by the hms genes (Hms-ECM). However, mutations in the Yptb BarA/UvrY/CsrB regulatory cascade enhance biofilm stability without dramatically increasing Hms-ECM production. We found that treatment with proteinase K enzyme effectively destabilized Yptb csrB mutant biofilms, suggesting that cell-cell interactions might be mediated by protein adhesins or extracellular matrix proteins. We identified an uncharacterized trimeric autotransporter lipoprotein (YPTB2394), repressed by csrB, which has been referred to as YadE. Biofilms made by a {Delta}yadE mutant strain were extremely sensitive to mechanical disruption. Overexpression of yadE in wild-type Yptb increased biofilm cohesion, similar to biofilms made by csrB or uvrY mutants. We found that the Rcs signaling cascade, which represses Hms-ECM production, activated expression of yadE. The yadE gene appears to be functional in Yptb but is a pseudogene in modern Y. pestis strains. Expression of functional yadE in Y. pestis KIM6+ altered the production of Hms-ECM and weakened biofilms made by these bacteria. This suggests that although the YadE autotransporter protein increases Yptb biofilm stability, it may be incompatible with Hms-ECM production that is essential for Y. pestis biofilm production in fleas. Inactivation of yadE in Y. pestis may be another instance of selective gene loss in the evolution of flea-borne transmission by this species. IMPORTANCEThe evolution of Yersinia pestis from its Y. pseudotuberculosis (Yptb) ancestor involved gene acquisition and gene losses, leading to differences in biofilm production. Characterizing the unique biofilm features of both species may provide better understanding of how each adapts to its specific niches. This study identifies a trimeric autotransporter YadE that promotes biofilm stability of Yptb but which has been inactivated in Y. pestis, likely because it is not compatible with Hms polysaccharide that is crucial for biofilms inside fleas. We also reveal that the Rcs signaling cascade, which represses Hms expression in Y. pestis, activates YadE in Yptb. The ability of Yptb to use polysaccharide or YadE protein for cell-cell adhesion may help it produce biofilms in different environments.