RRSP and RID Effector Domains Dominate the Virulence Impact of Vibrio vulnificus MARTX Toxin

Intragastric inoculation of mice with mutant Vibrio vulnificus reveals the impact of individual multifunctional autoprocessing repeats-in-toxins (MARTX) toxin effector domains on bacterial pathogenesis. Effector domains that disrupt Rho GTPase signaling exert the greatest effect on virulence, despite in vitro redundancy.The bacterial pathogen Vibrio vulnificus causes severe septic foodborne infections. The multifunctional autoprocessing repeats-in-toxins (MARTX) toxin is an important secreted virulence factor. The effector domain region is essential for lethal intestinal infection in mice, but the contribution of each of the 5 effector domains to infection has not been investigated. V. vulnificus mutants with varying effector domain content were inoculated intragastrically to mice, and the time to death was monitored to establish the contribution of each effector domain to overall virulence. Each strain was also tested for bacterial dissemination from the intestine to internal organs and for inhibition of phagocytosis. The effector domain region was required for V. vulnificus to inhibit phagocytosis by J774 macrophages, but no single effector domain was required. No single MARTX effector domain was necessary for bacterial dissemination. Nonetheless, overall survival of infected mice differed with respect to the infecting V. vulnificus strain. Removal of rid or rrsp significantly reduced the virulence potential of V. vulnificus, while deletion of duf1 or abh accelerated the time to death. Rho GTPases inactivation domain and Ras/Rap1-specific endopeptidase each exert greater effects on virulence than other MARTX domains, suggesting that modulation of the Rho/Ras family of GTPases is a critical function of the toxin during intestinal infection.