title: The Streptococcus mutans aminotransferase encoded by ilvE is regulated by CodY and CcpA.
abstract: The aminotransferase IlvE was implicated in the acid tolerance response of Streptococcus mutans when a mutation in its gene resulted in an acid-sensitive phenotype (B. Santiago, M. MacGilvray, R. C. Faustoferri, and R. G. Quivey, Jr., J. Bacteriol. 194:2010-2019, 2012). The phenotype suggested that amino acid metabolism is important for acid adaptation, as turnover of branched-chain amino acids (bcAAs) could provide important signals to modulate expression of genes involved in the adaptive process. Previous studies have demonstrated that ilvE is regulated in response to the external pH, though the mechanism is not yet established. CodY and CcpA have been shown to regulate expression of branched-chain amino acid biosynthetic genes, suggesting that the ability to sense carbon flow and the nutritional state of the cell also plays a role in the regulation of ilvE. Electrophoretic mobility shift assays using the ilvE promoter and a purified recombinant CodY protein provided evidence of the physical interaction between CodY and ilvE. In order to elucidate the signals that contribute to ilvE regulation, cat reporter fusions were utilized. Transcriptional assays demonstrated that bcAAs are signaling molecules involved in the repression of ilvE through regulation of CodY. In a codY deletion background, ilvE transcription was elevated, indicating that CodY acts a repressor of ilvE transcription. Conversely, in a ccpA deletion background, ilvE transcription was reduced, showing that CcpA activated ilvE transcription. The effects of both regulators were directly relevant for transcription of ilvE under conditions of acid stress, demonstrating that both regulators play a role in acid adaptation.
authors.affiliation: Center for Oral Biology, University of Rochester School of Medicine and Dentistry, Rochester, New York, USA.
authors.forename: Brendaliz
authors.initials: B
authors.lastname: Santiago
authors.forename: Maksym
authors.initials: M
authors.lastname: Marek
authors.forename: Roberta C
authors.initials: RC
authors.lastname: Faustoferri
authors.forename: Robert G
authors.initials: RG
authors.lastname: Quivey
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001431:Bacteriological Techniques; D021901:DNA, Intergenic / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D007532:Isoleucine; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013295:Streptococcus mutans / Q000235:genetics / Q000378:metabolism*; D000637:Transaminases / Q000235:genetics / Q000378:metabolism*; D014158:Transcription, Genetic
chemical_list: D001426:Bacterial Proteins; D021901:DNA, Intergenic; D011994:Recombinant Proteins; D007532:Isoleucine; D000637:Transaminases