title: Sialic acid transport and catabolism are cooperatively regulated by SiaR and CRP in nontypeable Haemophilus influenzae.
abstract: BACKGROUND
  The transport and catabolism of sialic acid, a critical virulence factor for nontypeable Haemophilus influenzae, is regulated by two transcription factors, SiaR and CRP.
  
  
  RESULTS
  Using a mutagenesis approach, glucosamine-6-phosphate (GlcN-6P) was identified as a co-activator for SiaR. Evidence for the cooperative regulation of both the sialic acid catabolic and transport operons suggested that cooperativity between SiaR and CRP is required for regulation. cAMP was unable to influence the expression of the catabolic operon in the absence of SiaR but was able to induce catabolic operon expression when both SiaR and GlcN-6P were present. Alteration of helical phasing supported this observation by uncoupling SiaR and CRP regulation. The insertion of one half-turn of DNA between the SiaR and CRP operators resulted in the loss of SiaR-mediated repression of the transport operon while eliminating cAMP-dependent induction of the catabolic operon when GlcN-6P was present. SiaR and CRP were found to bind to their respective operators simultaneously and GlcN-6P altered the interaction of SiaR with its operator.
  
  
  CONCLUSIONS
  These results suggest multiple novel features for the regulation of these two adjacent operons. SiaR functions as both a repressor and an activator and SiaR and CRP interact to regulate both operons from a single set of operators.
authors.affiliation: Department of Microbiology, Immunology, and Molecular Genetics, The University of Kentucky, Lexington, KY, USA. jason.johnston@uky.edu
authors.forename: Jason W
authors.initials: JW
authors.lastname: Johnston
authors.forename: Haider
authors.initials: H
authors.lastname: Shamsulddin
authors.forename: Anne-Frances
authors.initials: AF
authors.lastname: Miller
authors.forename: Michael A
authors.initials: MA
authors.lastname: Apicella
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D002373:Cyclic AMP Receptor Protein / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D006193:Haemophilus influenzae / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000096:biosynthesis*; D009875:Operator Regions, Genetic; D009876:Operon; D011485:Protein Binding
chemical_list: D001426:Bacterial Proteins; D002373:Cyclic AMP Receptor Protein; D019158:N-Acetylneuraminic Acid