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Microbiology (Reading) ; 151(Pt 1): 281-290, 2005 Jan.
Article in English | MEDLINE | ID: mdl-15632445

ABSTRACT

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.


Subject(s)
Aminoglycosides/biosynthesis , Bacterial Proteins/metabolism , DNA-Binding Proteins/metabolism , Streptomyces/metabolism , Up-Regulation , Bacterial Proteins/genetics , DNA-Binding Proteins/genetics , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression Regulation, Bacterial , Microscopy, Confocal , Molecular Sequence Data , Promoter Regions, Genetic , Recombinant Fusion Proteins/metabolism , Sequence Analysis, DNA , Streptomyces/genetics , Streptomyces/ultrastructure , Transcription, Genetic
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