Yersinia pseudotuberculosis blocks the phagosomal acidification of B10.A mouse macrophages through the inhibition of vacuolar H(+)-ATPase activity

Abstract

Yersinia pseudotuberculosis survived and multiplied in the phagosomes of B10.A mouse peritoneal macrophages. As one of the possible mechanisms for the bacteria's survival in the phagosomes, we demonstrated that live Y. pseudotuberculosis inhibited the phagosomal acidification; pH within phagosomes containing the live Y. pseudotuberculosis remained at about 6.0, whereas pH within phagosomes containing the dead Y. pseudotuberculosis fell to about 4. 5. This ability to inhibit intraphagosomal acidification was also shared by mutants lacking the 42 Md virulence plasmid, indicating that it is chromosomally encoded. The phagosomes containing dead bacteria raised the pH to 6.2 after the treatment of their macrophages with an inhibitor (bafilomycin A1) specific for V-ATPase. Although the amount of V-ATPase in the A and B subunits on the phagosomes was not significantly different between the live and dead bacteria infection, the phagosomes containing live bacteria had a 10-fold smaller V-ATPase activity than those containing the dead bacteria. These results indicated that the inhibition of phagosomal acidification by Y. pseudotuberculosis infection was due to the attenuation of V-ATPase activity, and not due to the exclusion of V-ATPase subunits from the phagosome membrane as found in Mycobacterium avium.