Although Shigella flexneri possesses the genes for two siderophore systems, enterobactin and aerobactin, the enterobactin system is only rarely utilized. To investigate the regulation of enterobactin expression in S. flexneri, all of the genes specifically required for synthesis and transport of enterobactin were cloned from both an expressing (Ent+) and a nonexpressing (Ent-) strain. Notable differences between the cloned genes included endonuclease restriction site changes and the presence of an IS1 element in the Ent- DNA. Southern hybridization revealed that this IS1 element, present at the 3' end of the entF gene, is conserved at this location in different strains and serotypes of Ent- S. flexneri. The Ent- cloned genes were tested for their ability to complement the defect in 11 different Escherichia coli enterobactin mutants. The Ent- genes fully complemented nine mutants but failed to complement the entF mutant AN117 and only partially complemented the entE mutant AN93. Whole-cell RNA isolated from E. coli and the Shigella strains was hybridized to 32P-labeled DNA containing the entB gene or a fragment carrying a portion of the entF gene. E. coli and the Ent+ Shigella strains exhibited derepression of transcription of these genes in low-iron media. Transcription in the Ent- strain remained repressed regardless of iron concentration. Expression of the entB and entF genes was also examined in an Ent- Shigella fur mutant. Expression of entF was only partially derepressed and entB remained fully repressed at all iron concentrations, suggesting that factors other than Fur are responsible for the repression of these enterobactin genes in the Ent- Shigella strains.
The ability of bacterial pathogens to acquire iron in the host is an essential component of the disease process. Pathogenic Enterobacteriaceae spp. may either scavenge host iron sources such as heme or induce high-affinity iron-transport systems to remove iron from host proteins. The ease with which iron is acquired from the host will be at least partially determined by the iron status of the host at the time of infection. In response to infection, mammalian hosts reduce serum iron levels and withhold iron from the invading microorganisms. Thus the competition for iron is an active process which influences the outcome of a host-bacterial interaction.
El Tor and non-O1 strains of Vibrio cholerae were analyzed to determine whether synthesis of secreted hemolysin was influenced by the concentration of iron in the medium. Synthesis of hemolysin was found to be iron regulated in both El Tor and non-O1 isolates. Increased levels of hemolytic activity were detected in supernatants of iron-starved cells. Spontaneous hemolysin-deficient mutants of one non-O1 strain were found to occur at high frequency. These variants also failed to synthesize vibriobactin, the iron transport compound utilized by V. cholerae. Another non-O1 strain was found to synthesize both hemolysin and vibriobactin constitutively. When the cloned Escherichia coli fur gene, encoded on the plasmid pABN203, was introduced into this constitutive strain, normal iron regulation of both hemolysin and vibriobactin was reestablished. The ability of V. cholerae to utilize mammalian iron compounds was determined, and it was found that both hemin and hemoglobin could serve as sole sources of iron.