caa a22 4500 605500568 CHVBK 20210128100556.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00253-015-6472-x doi (NATIONALLICENCE)springer-10.1007/s00253-015-6472-x Pathway-specific regulation revisited: cross-regulation of multiple disparate gene clusters by PAS-LuxR transcriptional regulators [Elektronische Daten] [Cláudia Vicente, Tamara Payero, Javier Santos-Aberturas, Eva Barreales, Antonio de Pedro, Jesús Aparicio] PAS-LuxR regulators are highly conserved proteins devoted to the control of antifungal production by binding to operators located in given promoters of polyene biosynthetic genes. The canonical operator of PimM, archetype of this class of regulators, has been used here to search for putative targets of orthologous protein PteF in the genome of Streptomyces avermitilis, finding 97 putative operators outside the pentaene filipin gene cluster (pte). The processes putatively affected included genetic information processing; energy, carbohydrate, and lipid metabolism; DNA replication and repair; morphological differentiation; secondary metabolite biosynthesis; and transcriptional regulation, among others. Seventeen of these operators were selected, and their binding to PimM DNA-binding domain was assessed by electrophoretic mobility shift assays. Strikingly, the protein bound all predicted operators suggesting a direct control over targeted processes. As a proof of concept, we studied the biosynthesis of the ATP-synthase inhibitor oligomycin whose gene cluster included two operators. Regulator mutants showed a severe loss of oligomycin production, whereas gene complementation of the mutant restored phenotype, and gene duplication in the wild-type strain boosted oligomycin production. Comparative gene expression analyses in parental and mutant strains by reverse transcription-quantitative polymerase chain reaction of selected olm genes corroborated production results. These results demonstrate that PteF is able to cross-regulate the biosynthesis of two related secondary metabolites, filipin and oligomycin, but might be extended to all the processes indicated above. This study highlights the complexity of the network of interactions in which PAS-LuxR regulators are involved and opens new possibilities for the manipulation of metabolite production in Streptomycetes. Springer-Verlag Berlin Heidelberg, 2015 Gene regulation nationallicence LuxR nationallicence Metabolic engineering nationallicence PAS domain nationallicence Polyene macrolide nationallicence Regulator binding site nationallicence Streptomyces nationallicence Vicente Cláudia Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut Payero Tamara Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut Santos-Aberturas Javier Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real, no. 1, 24006, León, Spain aut Barreales Eva Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut de Pedro Antonio Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut Aparicio Jesús Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/12(2015-06-01), 5123-5135 0175-7598 99:12<5123 2015 99 253 https://doi.org/10.1007/s00253-015-6472-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6472-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Vicente Cláudia Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut NATIONALLICENCE 700 1- Payero Tamara Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut NATIONALLICENCE 700 1- Santos-Aberturas Javier Institute of Biotechnology INBIOTEC, Parque Científico de León, Avda. Real, no. 1, 24006, León, Spain aut NATIONALLICENCE 700 1- Barreales Eva Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut NATIONALLICENCE 700 1- de Pedro Antonio Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut NATIONALLICENCE 700 1- Aparicio Jesús Area of Microbiology, Faculty of Biology, University of León, 24071, León, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/12(2015-06-01), 5123-5135 0175-7598 99:12<5123 2015 99 253