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The role of the  nfuA  gene encoding an iron-sulfur ([Fe-S]) cluster-delivery protein in the pathogenic bacterium  Pseudomonas aeruginosa  was investigated. The analysis of  nfuA  expression under various stress conditions showed that superoxide generators, a thiol-depleting agent and CuCl 2  highly induced  nfuA  expression. The expression of  nfuA  was regulated by a global [2Fe-2S] cluster containing the transcription regulator IscR. Increased expression of  nfuA  in the Δ iscR  mutant under uninduced conditions suggests that IscR acts as a transcriptional repressor. In vitro experiments revealed that IscR directly bound to a sequence homologous to the  Escherichia coli  Type-I IscR-binding motifs on a putative  nfuA  promoter that overlapped the -35 element. Binding of IscR prevented RNA polymerase from binding to the  nfuA  promoter, leading to repression of the  nfuA  transcription. Physiologically, deletion of  nfuA  reduced the bacterial ability to cope with oxidative stress, iron deprivation conditions and attenuated virulence in the  Caenorhabditis elegans  infection model. Site-directed mutagenesis analysis revealed that the conserved CXXC motif of the Nfu-type scaffold protein domain at the N-terminus was required for the NfuA functions in conferring the stress resistance phenotype. Furthermore, anaerobic growth of the Δ nfuA  mutant in the presence of nitrate was drastically retarded. This phenotype was associated with a reduction in the [Fe-S] cluster containing nitrate reductase enzyme activity. However, NfuA was not required for the maturation of [Fe-S]-containing proteins such as aconitase, succinate dehydrogenase, SoxR and IscR. Taken together, our results indicate that NfuA functions in [Fe-S] cluster delivery to selected target proteins that link to many physiological processes such as anaerobic growth, bacterial virulence and stress responses in  P .  aeruginosa .

plos one

Pseudomonas aeruginosa nfuA: Gene regulation and its physiological roles in sustaining growth under stress and anaerobic conditions and maintaining bacterial virulence