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Properties of Streptomyces albus J1074 mutant deficient in tRNALeu UAA gene bldA

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Abstract

Streptomyces albus J1074 is one of the most popular and convenient hosts for heterologous expression of gene clusters directing the biosynthesis of various natural metabolic products, such as antibiotics. This fuels interest in elucidation of genetic mechanisms that may limit secondary metabolism in J1074. Here, we report the generation and initial study of J1074 mutant, deficient in gene bldA for tRNALeu UAA, the only tRNA capable of decoding rare leucyl TTA codon in Streptomyces. The bldA deletion in J1074 resulted in a highly conditional Bld phenotype, with depleted formation of aerial hyphae on certain solid media. In addition, bldA mutant of J1074 was unable to produce endogenous antibacterial compounds and two heterologous antibiotics, moenomycin and aranciamycin, whose biosynthesis is directed by TTA-containing genes. We have employed a new TTA codon-specific β-galactosidase reporter system to provide genetic evidence that J1074 bldA mutant is impaired in translation of TTA. In addition, we have discussed the possible reasons for differences in the phenotypes of bldA mutants described here and in previous studies, providing knowledge to study bldA-based regulation of antibiotic biosynthesis.

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Acknowledgements

This work was supported by grant BG-41Nr from the Ministry of Education and Science of Ukraine (to B.O.).

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Authors and Affiliations

  1. Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 4 Hrushevskoho st., Lviv, 79005, Ukraine

    Oksana Koshla, Maria Lopatniuk, Ihor Rokytskyy, Oleksandr Yushchuk, Victor Fedorenko & Bohdan Ostash

  2. Actinobacteria Metabolic Engineering Group, Department of Pharmaceutical Biotechnology, Helmholtz Institute for Pharmaceutical Research Saarland, Saarland University, UdS Campus C2 3, 66123, Saarbrucken, Germany

    Maria Lopatniuk & Andriy Luzhetskyy

  3. Department of Physics of Earth, Ivan Franko National University of Lviv, 4 Hrushevskoho st., Lviv, 79005, Ukraine

    Yuriy Dacyuk

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  1. Oksana Koshla
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  2. Maria Lopatniuk
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Correspondence to Bohdan Ostash.

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Communicated by Erko Stackebrandt.

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Koshla, O., Lopatniuk, M., Rokytskyy, I. et al. Properties of Streptomyces albus J1074 mutant deficient in tRNALeu UAA gene bldA . Arch Microbiol 199, 1175–1183 (2017). https://doi.org/10.1007/s00203-017-1389-7

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