oneidensis Fur regulates genes AZD8931 supplier involved in iron homeostasis and acid resistance [10–13]. Consistently, many of these target genes have a recognizable “”Fur box”" in their promoters. In the present study, we further characterize a fur null mutant of S. oneidensis with regard to its ability to utilize succinate and fumarate. Unexpectedly,

HPLC analysis showed that the fur mutant was able to metabolize succinate and fumarate, and the growth of the mutant was enhanced in the presence of succinate and fumarate, indicating that the mutant can utilize these compounds. In addition, the expression of the TCA cycle genes acnA and sdhA was not down-regulated in the Selleck GW3965 mutant. These differences between S. oneidensis and E. coli were traced to the small RNA gene ryhB, which we identified Selleckchem Barasertib in several Shewanella species. Although S. oneidensis RyhB was up-regulated in the fur mutant, the TCA cycle genes did not appear to be regulated by RyhB. These results delineate differences in the gene regulation and physiological consequences of RyhB between S. oneidensis and E. coli. Results TCA cycle activity and regulation in the fur mutant We showed recently that S. oneidensis harboring a fur deletion in the genome was sensitive to acidic conditions and de-repressed genes encoding iron acquisition systems [11]. Similar observations

have been made in E. coli [14, 15], suggesting that the functional roles of Fur are conserved in these species. Since Fur acts as a pleiotropic transcription factor involved in multiple biological processes, we proceeded to examine its role in regulating TCA cycle enzymes. The involvement of Fur in this biological process has been established in E. coli and V. cholerae by observations that fur mutants are unable to grow in defined

media with succinate or fumarate as a carbon source [9, 16], and that genes encoding certain TCA cycle enzymes, such as succinate dehydrogenase (SdhABCD) and aconitase (AcnA), are significantly down-regulated in a fur mutant [7]. Our initial tests showed that neither succinate nor fumarate, when provided as the sole carbon source in M1 defined media, could support detectable growth of S. oneidensis type strain MR-1 (data not shown), making it unlikely to Morin Hydrate analyze the growth of MR-1 and fur null mutant. However, the complete set of TCA genes is present in S. oneidensis genome, and recent studies have shown that the bacterium is capable of metabolizing succinate and fumarate [17, 18]. To compare the metabolizing rates of the carbonates between MR-1 and the fur mutant, both strains were grown to mid-log phase with 10 mM lactate as the carbon source. Then equal numbers of cells (5 × 109) were washed and resuspended in fresh M1 medium with 10 mM lactate, succinate or fumarate as the sole carbon source.