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Microbiology 151 (2005), 281-290; DOI  10.1099/mic.0.27244-0
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Microbiology 151 (2005), 281-290; DOI  10.1099/mic.0.27244-0
© 2005 Society for General Microbiology

DNA-binding activity of LndI protein and temporal expression of the gene that upregulates landomycin E production in Streptomyces globisporus 1912

Yu. Rebets1, B. Ostash1, A. Luzhetskyy1,3, S. Kushnir1, M. Fukuhara2, A. Bechthold3, M. Nashimoto4, T. Nakamura2 and V. Fedorenko1

1 Department of Genetics and Biotechnology of Ivan Franko National University of L'viv, Grushevskogo st.4, L'viv 79005, Ukraine
2 Department of Microbiology, Niigata University of Pharmacy, Kami-Shinei-cho 5-13-2, Niigata 950-2081, Japan
3 Institut für Pharmazeutische Wissenschaften, Lehrstuhl für Pharmazeutische Biologie und Biotechnologie Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany
4 Department of Applied Life Science, Niigata University of Pharmacy and Applied Life Science, Kami-Shinei-cho 5-13-2, Niigata 950-2081, Japan

Correspondence
V. Fedorenko
v_fedorenko{at}franko.lviv.ua

The gene lndI is involved in the pathway-specific positive regulation of biosynthesis of the antitumour polyketide landomycin E in Streptomyces globisporus 1912. LndI was overexpressed in Escherichia coli as a protein C-terminally fused to the intein-chitin-binding-domain tag and purified in a one-step column procedure. Results of in vivo LndI titration, DNA gel mobility-shift assays and promoter-probing experiments indicate that LndI is an autoregulatory DNA-binding protein that binds to its own gene promoter and to the promoter of the structural gene lndE. Enhanced green fluorescent protein was used as a reporter to study the temporal and spatial pattern of lndI transcription. Expression of lndI started before cells entered mid-exponential phase and peak expression coincided with maximal accumulation of landomycin E and biomass. In solid-phase analysis, lndI expression was evident in substrate mycelia but was absent from aerial hyphae and spores.

Abbreviations: CBD, chitin-binding domain; EGFP, enhanced green fluorescent protein; EMSA, electrophoretic mobility-shift assay; LaE, landomycin E; PI, propidium iodide; SARP, Streptomyces antibiotic regulatory protein

The GenBank/EMBL/DDBJ accession number for the sequence reported in this paper is AY662671.




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R. Novakova, D. Homerova, L. Feckova, and J. Kormanec
Characterization of a regulatory gene essential for the production of the angucycline-like polyketide antibiotic auricin in Streptomyces aureofaciens CCM 3239
Microbiology, August 1, 2005; 151(8): 2693 - 2706.
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