DNA-binding activity of LndI protein and temporal expression of the gene that upregulates landomycin E production in Streptomyces globisporus 1912.
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.
Rebets
Yu
Y
Department of Genetics and Biotechnology of Ivan Franko National University of L'viv, Grushevskogo st.4, L'viv 79005, Ukraine.
Ostash
B
B
Department of Genetics and Biotechnology of Ivan Franko National University of L'viv, Grushevskogo st.4, L'viv 79005, Ukraine.
Luzhetskyy
A
A
Institut für Pharmazeutische Wissenschaften, Lehrstuhl für Pharmazeutische Biologie und Biotechnologie Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany.|Department of Genetics and Biotechnology of Ivan Franko National University of L'viv, Grushevskogo st.4, L'viv 79005, Ukraine.
Kushnir
S
S
Department of Genetics and Biotechnology of Ivan Franko National University of L'viv, Grushevskogo st.4, L'viv 79005, Ukraine.
Fukuhara
M
M
Department of Microbiology, Niigata University of Pharmacy, Kami-Shinei-cho 5-13-2, Niigata 950-2081, Japan.
Bechthold
A
A
Institut für Pharmazeutische Wissenschaften, Lehrstuhl für Pharmazeutische Biologie und Biotechnologie Albert-Ludwigs-Universität Freiburg, Stefan-Meier-Strasse 19, 79104 Freiburg, Germany.
Nashimoto
M
M
Department of Applied Life Science, Niigata University of Pharmacy and Applied Life Science, Kami-Shinei-cho 5-13-2, Niigata 950-2081, Japan.
Nakamura
T
T
Department of Microbiology, Niigata University of Pharmacy, Kami-Shinei-cho 5-13-2, Niigata 950-2081, Japan.
Fedorenko
V
V
Department of Genetics and Biotechnology of Ivan Franko National University of L'viv, Grushevskogo st.4, L'viv 79005, Ukraine.
D000617:Aminoglycosides / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure; D014158:Transcription, Genetic; D015854:Up-Regulation
D000617:Aminoglycosides; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D011993:Recombinant Fusion Proteins; C113593:landomycin E