Identification of Novel Host Interactors of Effectors Secreted by <i>Salmonella</i> and <i>Citrobacter</i>

Abstract

Many pathogenic bacteria of the family <i>Enterobacteriaceae</i> use type III secretion systems to inject virulence proteins, termed "effectors," into the host cell cytosol. Although host-cellular activities of several effectors have been demonstrated, the function and host-targeted pathways of most of the effectors identified to date are largely undetermined. To gain insight into host proteins targeted by bacterial effectors, we performed coaffinity purification of host proteins from cell lysates using recombinant effectors from the <i>Enterobacteriaceae</i> intracellular pathogens <i>Salmonella enterica</i> serovar Typhimurium and <i>Citrobacter rodentium</i>. We identified 54 high-confidence host interactors for the <i>Salmonella</i> effectors GogA, GtgA, GtgE, SpvC, SrfH, SseL, SspH1, and SssB collectively and 21 interactors for the <i>Citrobacter</i> effectors EspT, NleA, NleG1, and NleK. We biochemically validated the interaction between the SrfH <i>Salmonella</i> protein and the extracellular signal-regulated kinase 2 (ERK2) host protein kinase, which revealed a role for this effector in regulating phosphorylation levels of this enzyme, which plays a central role in signal transduction. <b>IMPORTANCE</b> During infection, pathogenic bacteria face an adverse environment of factors driven by both cellular and humoral defense mechanisms. To help evade the immune response and ultimately proliferate inside the host, many bacteria evolved specialized secretion systems to deliver effector proteins directly into host cells. Translocated effector proteins function to subvert host defense mechanisms. Numerous pathogenic bacteria use a specialized secretion system called type III secretion to deliver effectors into the host cell cytosol. Here, we identified 75 new host targets of <i>Salmonella</i> and <i>Citrobacter</i> effectors, which will help elucidate their mechanisms of action.