title: Biosynthesis of the Fluorinated Natural Product Nucleocidin in Streptomyces calvus Is Dependent on the bldA-Specified Leu-tRNA(UUA) Molecule.
abstract: Nucleocidin is one of the very few natural products known to contain fluorine. Mysteriously, the nucleocidin producer Streptomyces calvus ATCC 13382 has not been observed to synthesize the compound since its discovery in 1956. Here, we report that complementation of S. calvus ATCC 13382 with a functional bldA-encoded Leu-tRNA(UUA) molecule restores the production of nucleocidin. Nucleocidin was detected in culture extracts by (19) F NMR spectroscopy, HPLC-ESI-MS, and HPLC-continuum source molecular absorption spectroscopy for fluorine-specific detection. The molecule was purified from a large-scale culture and definitively characterized by NMR spectroscopy and high-resolution MS. The nucleocidin biosynthetic gene cluster was identified by the presence of genes encoding the 5'-O-sulfamate moiety and confirmed by gene disruption. Two of the genes within the nucleocidin biosynthetic gene cluster contain TTA codons, thus explaining the dependence on bldA and resolving a 60-year-old mystery.
authors.affiliation: Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada.
authors.forename: Xi Ming
authors.initials: XM
authors.lastname: Zhu
authors.affiliation: Department of Pharmaceutical Biology and Biotechnology, Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104, Freiburg, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Hackl
authors.affiliation: Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1200 Main Street West, Hamilton, Ontario, L8N 3Z5, Canada.
authors.forename: Maulik N
authors.initials: MN
authors.lastname: Thaker
authors.affiliation: Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1200 Main Street West, Hamilton, Ontario, L8N 3Z5, Canada.
authors.forename: Lindsay
authors.initials: L
authors.lastname: Kalan
authors.affiliation: Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen, AB24 3UE, UK.
authors.forename: Claudia
authors.initials: C
authors.lastname: Weber
authors.affiliation: Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen, AB24 3UE, UK.
authors.forename: Dagmar S
authors.initials: DS
authors.lastname: Urgast
authors.affiliation: Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen, AB24 3UE, UK.
authors.forename: Eva M
authors.initials: EM
authors.lastname: Krupp
authors.affiliation: Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada.
authors.forename: Alyssa
authors.initials: A
authors.lastname: Brewer
authors.affiliation: Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Vanner
authors.affiliation: Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada.
authors.forename: Anjuli
authors.initials: A
authors.lastname: Szawiola
authors.affiliation: Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1200 Main Street West, Hamilton, Ontario, L8N 3Z5, Canada.
authors.forename: Grace
authors.initials: G
authors.lastname: Yim
authors.affiliation: Trace Element Speciation Laboratory, Department of Chemistry, University of Aberdeen, Aberdeen, AB24 3UE, UK.
authors.forename: Jörg
authors.initials: J
authors.lastname: Feldmann
authors.affiliation: Department of Pharmaceutical Biology and Biotechnology, Institute of Pharmaceutical Sciences, Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104, Freiburg, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bechthold
authors.affiliation: Michael G. DeGroote Institute for Infectious Disease Research, McMaster University, 1200 Main Street West, Hamilton, Ontario, L8N 3Z5, Canada.
authors.forename: Gerard D
authors.initials: GD
authors.lastname: Wright
authors.affiliation: Department of Chemistry, Queen's University, Chernoff Hall, 90 Bader Lane, Kingston, Ontario, K7L 3N6, Canada. dlzechel@chem.queensu.ca.
authors.forename: David L
authors.initials: DL
authors.lastname: Zechel
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives* / Q000032:analysis / Q000096:biosynthesis / Q000737:chemistry; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001688:Biological Products / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D005461:Fluorine / Q000737:chemistry; D054879:Halogenation; D013058:Mass Spectrometry; D005810:Multigene Family; D016366:Open Reading Frames / Q000235:genetics; D011683:Purine-Nucleoside Phosphorylase / Q000235:genetics / Q000378:metabolism; D012356:RNA, Transfer, Leu / Q000235:genetics / Q000378:metabolism*; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D001688:Biological Products; D012356:RNA, Transfer, Leu; D005461:Fluorine; D011683:Purine-Nucleoside Phosphorylase; C005318:nucleocidin; D000241:Adenosine
keywords: Streptomyces calvus; bldA; fluorine; nucleocidin; sulfamate