Proteomic analysis reveals the wide-ranging effects of the novel, iron-responsive regulator RirA in Rhizobium leguminosarum bv. viciae

Abstract

The wide-ranging effects of RirA, a novel Fe-responsive regulator of gene expression in Rhizobium leguminosarum bv. viciae, were monitored on 2D gels. Approximately 100 proteins were expressed at higher levels in a RirA(-) mutant, compared to wild type. These included the products of the sufS(2)BCDS(1)XA operon, which probably specifies the synthesis of [FeS] clusters. Using lac fusions, this operon was confirmed to be regulated by RirA in response to Fe availability. Genes for some ABC transporters, and a protein that may be involved in making a phenazine-like molecule, were also repressed by Fe in a RirA-dependent way. Strikingly, at least 17 proteins were reduced in abundance in the RirA(-) mutant. These included three ABC transporters, a GatB-like enzyme involved in tRNA modification, and a protein that may confer bacteriocin resistance. As judged by lac reporter fusions, this apparently positive control by RirA was probably due to post-transcriptional effects, in at least some cases. Therefore, although RirA shows no sequence similarity to Fur or DtxR, it functions as a wide-ranging, Fe-responsive regulator.