Analysis of Mutations of the Strains of Streptomyces fradiae ATCC 19609-Olg2R Resistant to (33S)-Azido-33-Deoxyoligomycin A

Resumo

In the present work, we analyze the mutant strains of Streptomyces fradiae ATCC 19609 resistant to (33S)-azido-33-deoxyoligomycin A, obtained at a frequency of 10^–8 to 2.5 × 10^–9. Analysis of the obtained mutant strains allows them to be divided into two groups based on the level of resistance to oligomycin A. One mutant out of 15 obtained is resistant to oligomycin A at MIC of 0.1 nm/mL, and 14 mutant strains are resistant at MIC of 0.01 nm/mL of oligomycin A (MIC for the initial strain: 0.001 nm/mL). Using PCR analysis and subsequent sequencing by the Sanger method, three candidate genes the products of which are related to resistance to oligomycin A are analyzed: helicase IV, the C subunit and the A subunit of FoF1-ATP synthase. It is found that, in the genomes of all 15 mutants, there is a single nucleotide substitution (SNP) in the helicase IV gene, while in one strain most resistant to oligomycin A there is an additional SNP in the gene of the A subunit of FoF1-ATP synthase. This mutant strain will be used to obtain the mutants of resistance to oligomycin A with the aim of further sequencing of genomes and detection of biotarget of oligomycin A.