Cloning of the netropsin resistance genes from Streptomyces flavopersicus NRRL 2820.
Streptomyces flavopersicus NRRL 2820 (synonym: Streptomyces netropsis DSM40093) is resistant to the N-methylpyrrole-containing oligopeptide antibiotic netropsin. A 9.38 kb DNA-fragment was isolated from a genomic library of Streptomyces flavopersicus using an Escherichia coli-Streptomyces lividans shuttle vector which enables S. lividans to grow on netropsin-containing agar plates. By subcloning, the resistance was conferred to a 5.9 kb Eco RV fragment. DNA sequence analysis of this Eco RV fragment revealed two open reading frames (netP1 , 1556 bp and netP2 , 1773 bp). The deduced proteins share significant similarity to each other (27% identity) and to the large family to ABC-type multidrug resistance proteins. In each protein a conserved transmembrane and ATP binding domain was identified. Deletion analysis showed that both proteins are necessary for netropsin resistance indicating that the proteins form a heterodimeric ABC-transporter exporting netropsin.
Stumpp
Tina
T
Institut für Industrielle Genetik, Universität Stuttgart, Allmandring 31, 70569 Stuttgart, Germany.
Himbert
Sarah
S
Altenbuchner
Josef
J
D000255:Adenosine Triphosphate / Q000378:metabolism; D004269:DNA, Bacterial; D004351:Drug Resistance / Q000235:genetics; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009429:Netropsin / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D016366:Open Reading Frames; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013302:Streptomyces / Q000187:drug effects* / Q000235:genetics*
D004269:DNA, Bacterial; D009429:Netropsin; D000255:Adenosine Triphosphate