title: The LysR-type transcriptional regulator (LTTR) AlsR indirectly regulates expression of the Bacillus subtilis bdhA gene encoding 2,3-butanediol dehydrogenase.
abstract: Bacillus subtilis ferments pyruvate to 2,3-butanediol via α-acetolactate synthase, α-acetolactate decarboxylase, and butanediol dehydrogenase (BDH), encoded by the alsSD operon and the unlinked monocistronic bdhA gene, respectively. Upstream and divergent from alsSD is the alsR gene that encodes AlsR, a member of the LysR-type transcriptional regulator family. AlsR directly stimulates alsSD transcription by binding to characteristic sites preceding the alsS promoter, but its effect on bdhA expression was unknown. The effect of AlsR on bdhA expression was assessed in a wild-type strain and a congenic strain carrying an alsR::spc knockout mutation by measuring: (a) expression of a transcriptional bdhA-lacZ fusion; (b) bdhA mRNA steady-state levels by quantitative reverse transcriptase PCR; and (c) expression of BDH enzymatic activity. Activation of bdhA expression occurred in early stationary phase, and expression was lowered, but not abolished, in the alsR::spc mutant. Mapping the transcriptional start site of bdhA by primer extension revealed a 268-nucleotide 5'-untranslated region preceding the bdhA initiation methionine codon. Transcription initiation was not reduced in the alsR::spc mutant, and by electrophoretic mobility shift assay, purified AlsR protein did not bind to the bdhA promoter region, suggesting that bdhA expression is indirectly under AlsR transcriptional control.
authors.affiliation: Department of Microbiology and Cell Science, University of Florida, Gainesville, FL, USA.
authors.forename: Rafael R
authors.initials: RR
authors.lastname: de Oliveira
authors.forename: Wayne L
authors.initials: WL
authors.lastname: Nicholson
mesh_terms: D020121:5' Untranslated Regions; D000429:Alcohol Oxidoreductases / Q000096:biosynthesis* / Q000235:genetics; D019677:Artificial Gene Fusion; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D002072:Butylene Glycols / Q000378:metabolism*; D018387:Codon, Initiator; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D055786:Gene Knockout Techniques; D017930:Genes, Reporter; D019289:Pyruvic Acid / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
chemical_list: D020121:5' Untranslated Regions; D002072:Butylene Glycols; D018387:Codon, Initiator; D014157:Transcription Factors; C026978:2,3-butylene glycol; D019289:Pyruvic Acid; D000429:Alcohol Oxidoreductases; C019801:butanediol dehydrogenase; D001616:beta-Galactosidase