title: The DyP-type peroxidase DtpA is a Tat-substrate required for GlxA maturation and morphogenesis in Streptomyces.
abstract: The filamentous bacterium Streptomyces lividans depends on the radical copper oxidase GlxA for the formation of reproductive aerial structures and, in liquid environments, for the formation of pellets. Incorporation of copper into the active site is essential for the formation of a cross-linked tyrosyl-cysteine cofactor, which is needed for enzymatic activity. In this study, we show a crucial link between GlxA maturation and a group of copper-related proteins including the chaperone Sco and a novel DyP-type peroxidase hereinafter called DtpA. Under copper-limiting conditions, the sco and dtpA deletion mutants are blocked in aerial growth and pellet formation, similarly to a glxA mutant. Western blot analysis showed that GlxA maturation is perturbed in the sco and dtpA mutants, but both maturation and morphology can by rescued by increasing the bioavailability of copper. DtpA acts as a peroxidase in the presence of GlxA and is a substrate for the twin-arginine translocation (Tat) translocation pathway. In agreement, the maturation status of GlxA is also perturbed in tat mutants, which can be compensated for by the addition of copper, thereby partially restoring their morphological defects. Our data support a model wherein a copper-trafficking pathway and Tat-dependent secretion of DtpA link to the GlxA-dependent morphogenesis pathway.
authors.affiliation: Molecular Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands.
authors.forename: Marloes L C
authors.initials: ML
authors.lastname: Petrus
authors.affiliation: Molecular Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands.
authors.forename: Erik
authors.identifier: http://orcid.org/0000-0002-5059-069X
authors.initials: E
authors.lastname: Vijgenboom
authors.affiliation: School of Biological Science, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK.
authors.forename: Amanda K
authors.initials: AK
authors.lastname: Chaplin
authors.affiliation: School of Biological Science, University of Essex, Wivenhoe Park, Colchester CO4 3SQ, UK.
authors.forename: Jonathan A R
authors.initials: JA
authors.lastname: Worrall
authors.affiliation: Molecular Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands.
authors.forename: Gilles P
authors.initials: GP
authors.lastname: van Wezel
authors.affiliation: Molecular Biotechnology, Institute of Biology, Leiden University, Sylviusweg 72, 2333 BE Leiden, The Netherlands d.claessen@biology.leidenuniv.nl.
authors.forename: Dennis
authors.identifier: http://orcid.org/0000-0002-0789-2633
authors.initials: D
authors.lastname: Claessen
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D003300:Copper / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005798:Genes, Bacterial; D015151:Immunoblotting; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009195:Peroxidase / Q000378:metabolism*; D010641:Phenotype; D048170:Streptomyces lividans / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D003300:Copper; D009195:Peroxidase
keywords: DyP-type peroxidase; GlxA maturation; cuproenzyme; glycan; morphogenesis; twin-arginine translocation