title: The effect of specific rhlA-las-box mutations on DNA binding and gene activation by Pseudomonas aeruginosa quorum-sensing transcriptional regulators RhlR and LasR.
abstract: Pseudomonas aeruginosa is a free-living bacterium and an important opportunistic pathogen. The genes coding for virulence-associated traits are regulated at the level of transcription by the quorum-sensing response. In this response, the regulator LasR coupled with the autoinducer 3-oxo-dodecanoyl homoserine lactone (3O-C12-HSL) activates transcription of genes for several virulence factors. LasR/3O-C12-HSL also activates transcription of rhlR, the gene coding for the transcriptional regulator RhlR, and of rhlI that encodes the synthase that produces the autoinducer butanoyl-homoserine lactone (C4-HSL) that interacts with RhlR. Genes activated by RhlR/C4-HSL include those involved in rhamnolipids production (like the rhlAB operon) and lecA, coding for PA-I lectin. The molecular basis of LasR/3O-C12-HSL- and RhlR/C4-HSLDNA-binding specificity (at the so-called las-boxes) has not been clearly determined, and the aim of this work was to contribute to its understanding. Therefore, we analyzed the interaction of LasR and RhlR to variants of the rhlA-las-box that were constructed based on the comparison of this las-box to the las-box of lecA. We conclude that LasR and RhlR DNA-binding specificity is a complex multifactorial phenomenon in which both positive and negative effects are involved and that binding of these proteins does not necessarily result in gene activation.
authors.affiliation: Departamento de Biología Molecular y Biotecnología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, México, México.
authors.forename: Abigail
authors.initials: A
authors.lastname: González-Valdez
authors.forename: Luis
authors.initials: L
authors.lastname: Servín-González
authors.forename: Katy
authors.initials: K
authors.lastname: Juárez
authors.forename: Alberto
authors.initials: A
authors.lastname: Hernandez-Aligio
authors.forename: Gloria
authors.initials: G
authors.lastname: Soberón-Chávez
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial / Q000378:metabolism; D009154:Mutation; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000502:physiology*; D053038:Quorum Sensing; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism*; D015533:Transcriptional Activation
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; C080365:LasR protein, Pseudomonas aeruginosa; C086515:RhlR protein, Pseudomonas aeruginosa; D015534:Trans-Activators
keywords: consensus-sequences; las-box-mutations; protein-DNA-interaction