title: Regulation of the Escherichia coli allantoin regulon: coordinated function of the repressor AllR and the activator AllS.
abstract: The allantoin regulon of Escherichia coli, formed by three operons expressed from promoters allA(P), gcl(P) and allD(P), is involved in the anaerobic utilization of allantoin as nitrogen source. The expression of these operons is under the control of the repressor AllR. The hyperinduction of one of these promoters (allD(P)) by allantoin in an AllR defective mutant suggested the action of another regulator, presumably of activator type. In this work we have identified ybbS (proposed gene name allS), divergently transcribed from allA, as the gene encoding this activator. Analysis of the expression of the three structural operons in DeltaallS mutant showed that the expression from allD(P) was abolished, suggesting that AllS is essential for the expression of the corresponding operon. In a wild-type strain expression of allS takes place mainly anaerobically and is hyperinduced when the nitrogen source limits growth. However, expression of allS is independent of regulators of the Ntr response, NtrC or Nac. Band shift experiments showed that AllR binds to DNA containing the allS-allA intergenic region and the gcl(P) promoter and its binding is abolished by glyoxylate. Both DNA fragments contain a highly conserved inverted repeat, which after site-directed mutagenesis, has been proven to be the AllR-binding site. This site displays similarity with the IclR family recognized consensus. Interaction of AllR with the single operator present in the allS-allA intergenic region prevented binding of RNA polymerase to either of the two divergent promoters. The regulator AllS interacts only with allD(P) even in the absence of allantoin. Analysis of this promoter allowed us to identify an inverted repeat as a motif for AllS binding. We propose a model for the coordinate control of the allantoin regulon by AllR and AllS.
authors.affiliation: Department of Biochemistry, Faculty of Pharmacy, University of Barcelona, Avda. Diagonal 643, 08028 Barcelona, Spain.
authors.forename: Maria R
authors.initials: MR
authors.lastname: Rintoul
authors.forename: Eva
authors.initials: E
authors.lastname: Cusa
authors.forename: Laura
authors.initials: L
authors.lastname: Baldomà
authors.forename: Josefa
authors.initials: J
authors.lastname: Badia
authors.forename: Larry
authors.initials: L
authors.lastname: Reitzer
authors.forename: Juan
authors.initials: J
authors.lastname: Aguilar
mesh_terms: D000481:Allantoin / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002874:Chromosome Mapping; D016384:Consensus Sequence; D004268:DNA-Binding Proteins / Q000235:genetics; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008957:Models, Genetic; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009584:Nitrogen / Q000378:metabolism; D009876:Operon; D011401:Promoter Regions, Genetic; D018085:Regulon; D012097:Repressor Proteins / Q000235:genetics / Q000502:physiology*; D016415:Sequence Alignment; D017384:Sequence Deletion; D015534:Trans-Activators / Q000235:genetics / Q000502:physiology*; D014158:Transcription, Genetic; D015533:Transcriptional Activation
chemical_list: D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D000481:Allantoin; D012321:DNA-Directed RNA Polymerases; D009584:Nitrogen