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Enteropathogenic and enterohaemorrhagic  Escherichia coli  express a cell cycle-inhibiting factor (Cif), that is injected into host cells via a Type III secretion system (T3SS) leading to arrest of cell division, delayed apoptosis and cytoskeletal rearrangements. A homologue of Cif has been identified in  Burkholderia pseudomallei  (CHBP;  C if  h omologue in   B .  p seudomallei ; BPSS1385), which shares catalytic activity, but its prevalence, secretion and function are ill-defined. Among 43 available  B. pseudomallei  genome sequences, 33 genomes (76.7%) harbor the gene encoding CHBP. Western blot analysis using antiserum raised to a synthetic CHBP peptide detected CHBP in 46.6% (7/15) of clinical  B. pseudomallei  isolates from the endemic area. Secretion of CHBP into bacterial culture supernatant could not be detected under conditions where a known effector (BopE) was secreted in a manner dependent on the Bsa T3SS. In contrast, CHBP could be detected in U937 cells infected with  B. pseudomallei  by immunofluorescence microscopy and Western blotting in a manner dependent on  bsaQ . Unlike  E. coli  Cif, CHBP was localized within the cytoplasm of  B. pseudomallei -infected cells. A  B. pseudomallei chbP  insertion mutant showed a significant reduction in cytotoxicity and plaque formation compared to the wild-type strain that could be restored by plasmid-mediated  trans -complementation. However, there was no defect in actin-based motility or multinucleated giant cell formation by the  chbP  mutant. The data suggest that the level or timing of CHBP secretion differs from a known Bsa-secreted effector and that CHBP is required for selected virulence-associated phenotypes  in vitro .

Analysis of the Prevalence, Secretion and Function of a Cell Cycle-Inhibiting Factor in the Melioidosis Pathogen Burkholderia pseudomallei