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Insights into the Biological Properties of Ligands and Identity of Operator Site for LanK Protein Involved in Landomycin Production

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Abstract

LanK is a TetR type regulatory protein that coordinates the late steps of the biosynthesis of the landomycin family of antitumor angucyclic polyketides and their export from the cells of Streptomyces cyanogenus S136. We recently described the structure of LanK and showed that it is the carbohydrate portion of the landomycins that is responsible for abrogating the repressing effect of LanK on landomycin production and export. The effect has been established in a series of in vitro tests using synthetic analogs of the landomycin carbohydrate chains. Whether such synthetic compounds would function as effector molecules for LanK under in vivo conditions remained unknown. Furthermore, the location and identity of LanK operator sites within the lanK-lanJ intergenic region (lanKJp) was unknown. Here we report that methoxyphenyl analogs of tri- and hexasaccharide chains of landomycins cannot function as LanK ligands when applied externally to the reporter strain. The lability of these compounds to cellular media and/or their poor penetration into the cells could explain our observations. The LanK operator site has been mapped to a 14-bp region of lanKJp that includes a plausible -35 site upstream of the lanK start codon in a series of electrophoretic DNA mobility shift assays. This opens the door to studies of the DNA-LanK interaction at a single nucleotide resolution level.

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All the data are present in the manuscript and Supplementary Materials. Original versions of all images will be provided by the corresponding author upon reasonable request.

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Funding

The work was supported by grant BG-21F 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

    Maksym Deneka, Iryna Ostash & Bohdan Ostash

  2. Department of Chemistry, Tufts University, 62 Talbot Ave, Medford, MA, 02155, USA

    Subbarao Yalamanchili & Clay S. Bennett

  3. Department of Genetics and Biotechnology, Ivan Franko National University of Lviv, 4 Hrushevskoho St.,Rm 102, Lviv, 79005, Ukraine

    Bohdan Ostash

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  1. Maksym Deneka
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  2. Iryna Ostash
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  3. Subbarao Yalamanchili
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  4. Clay S. Bennett
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  5. Bohdan Ostash
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Contributions

Conceptualization: BO, Methodology: IO, SY, Formal analysis and investigation: MD, BO, Writing—original draft preparation: BO, MD, Writing—review and editing: BO, CB, Funding acquisition: BO, Resources: SY, IO, CB, Supervision: BO.

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Correspondence to Bohdan Ostash.

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Deneka, M., Ostash, I., Yalamanchili, S. et al. Insights into the Biological Properties of Ligands and Identity of Operator Site for LanK Protein Involved in Landomycin Production. Curr Microbiol 81, 5 (2024). https://doi.org/10.1007/s00284-023-03528-1

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