title: Characterisation by molecular cloning of two genes from Streptomyces verticillus encoding resistance to bleomycin.
abstract: Extracts of a bleomycin (Bm)-producing Streptomyces verticillus ATCC15003 were found to possess an acetyltransferase activity which inactivates Bm in the presence of acetyl coenzyme A. DNA fragments of S. verticillus were introduced into S. lividans by cloning and transformants selected for resistance to Bm. Deletion mapping and subcloning of a 6-kb DNA fragment showed the presence of two resistance determinants, blmA and blmB. The acetyltransferase activity was encoded by blmB; nucleotide sequence analysis identified an ORF consisting of 301 amino acids (aa) proposed to be that of Bm acetyltransferase (Bat). S. lividans and Escherichia coli transformants harboring plasmids carrying blmB produced an acetyltransferase which modified and determined resistance to Bm and structurally related antibiotics; this resistance gene has potential as a selective marker in gene transfer studies. Nucleotide sequence analysis of blmA revealed an ORF encoding 122 aa that had significant sequence similarity to the gene encoding the Bm-binding protein (Shble) identified by Gatignol et al. [FEBS Lett. 230 (1988) 171-175] in Streptoalloteichus hindustanus, the tallysomycin-producer. The blmA gene was expressed in E. coli and the resulting protein, like the Shble protein, prevents in vitro Bm-induced DNA breakage.
authors.affiliation: Unité de Génie Microbiologique, Institut Pasteur, Paris, France.
authors.forename: M
authors.initials: M
authors.lastname: Sugiyama
authors.forename: C J
authors.initials: CJ
authors.lastname: Thompson
authors.forename: T
authors.initials: T
authors.lastname: Kumagai
authors.forename: K
authors.initials: K
authors.lastname: Suzuki
authors.forename: R
authors.initials: R
authors.lastname: Deblaere
authors.forename: R
authors.initials: R
authors.lastname: Villarroel
authors.forename: J
authors.initials: J
authors.lastname: Davies
mesh_terms: D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D001761:Bleomycin / Q000494:pharmacology*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics*; D017353:Gene Deletion; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
chemical_list: D004269:DNA, Bacterial; D001761:Bleomycin; D000123:Acetyltransferases