title: The AraC-type regulator RipA represses aconitase and other iron proteins from Corynebacterium under iron limitation and is itself repressed by DtxR.
abstract: The mRNA level of the aconitase gene acn of Corynebacterium glutamicum is reduced under iron limitation. Here we show that an AraC-type regulator, termed RipA for "regulator of iron proteins A," is involved in this type of regulation. A C. glutamicum DeltaripA mutant has a 2-fold higher aconitase activity than the wild type under iron limitation, but not under iron excess. Comparison of the mRNA profiles of the DeltaripA mutant and the wild type revealed that the acn mRNA level was increased in the DeltaripA mutant under iron limitation, but not under iron excess, indicating a repressor function of RipA. Besides acn, some other genes showed increased mRNA levels in the DeltaripA mutant under iron starvation (i.e. those encoding succinate dehydrogenase (sdhCAB), nitrate/nitrite transporter and nitrate reductase (narKGHJI), isopropylmalate dehydratase (leuCD), catechol 1,2-dioxygenase (catA), and phosphotransacetylase (pta)). Most of these proteins contain iron. Purified RipA binds to the upstream regions of all operons mentioned above and in addition to that of the catalase gene (katA). From 13 identified binding sites, the RipA consensus binding motif RRGCGN(4)RYGAC was deduced. Expression of ripA itself is repressed under iron excess by DtxR, since purified DtxR binds to a well conserved binding site upstream of ripA. Thus, repression of acn and the other target genes indicated above under iron limitation involves a regulatory cascade of two repressors, DtxR and its target RipA. The modulation of the intracellular iron usage by RipA supplements mechanisms for iron acquisition that are directly regulated by DtxR.
authors.affiliation: Institut für Biotechnologie 1, Forschungszentrum Jülich, Jülich D-52425, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Wennerhold
authors.forename: Andreas
authors.initials: A
authors.lastname: Krug
authors.forename: Michael
authors.initials: M
authors.lastname: Bott
mesh_terms: D000154:Aconitate Hydratase / Q000235:genetics* / Q000378:metabolism; D051777:AraC Transcription Factor / Q000235:genetics / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000494:pharmacology; D001665:Binding Sites; D002374:Catalase / Q000235:genetics; D048230:Corynebacterium glutamicum / Q000737:chemistry* / Q000201:enzymology; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000494:pharmacology; D004926:Escherichia coli; D005786:Gene Expression Regulation / Q000187:drug effects*; D007501:Iron / Q000494:pharmacology; D009154:Mutation; D019159:Nonheme Iron Proteins / Q000235:genetics*; D009876:Operon; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D011994:Recombinant Proteins
chemical_list: D051777:AraC Transcription Factor; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C075586:DtxR protein, Corynebacterium diphtheriae; D019159:Nonheme Iron Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins; D004247:DNA; D007501:Iron; D002374:Catalase; D000154:Aconitate Hydratase