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Gene ; 425(1-2): 9-16, 2008 Dec 01.
Article in English | MEDLINE | ID: mdl-18761063

ABSTRACT

The gamma-butyrolactone-autoregulator signalling system is widely distributed across many Streptomyces species and it controls secondary metabolism and/or morphological differentiation. IM-2 [(2R,3R,1'R)-2-1'-hydroxybutyl-3-hydroxymethyl-gamma-butanolide] is a gamma-butyrolactone autoregulator which, in Streptomyces lavendulae FRI-5, switches off the production of D-cycloserine, but switches on the production of several nucleoside antibiotics and blue pigment. In the IM-2 system, an IM-2 specific receptor (FarA) plays a critical role in the biosynthetic regulation of these metabolites, including IM-2 itself. Here, we identified five additional regulatory genes in the farA-flanking region and demonstrated that, in addition to farA, at least two more genes (farR1 and farR2) are involved in the IM-2/FarA system as the direct transcriptional target of FarA. The gel-shift assay revealed that FarA was bound to the upstream region of the four genes (including farR1 and farR2) in an IM-2-dependent manner. The FarA-binding sites were localized by DNase I footprinting to 27- to 33-bp palindromic structures, suggesting that FarA-binding sequences consist of two conserved hexamers separated by six nucleotides. Both farR1 and farR2 were transcribed in a growth-dependent manner, and marked expression was induced in the presence of IM-2, whereas transcripts of other two genes were not detected under the cultivation conditions used. The FarA-binding sites of farR1 and far2 overlap the promoter regions, suggesting that FarA represses the transcription of these two genes in the absence of IM-2 by inhibiting RNA polymerase access.


Subject(s)
4-Butyrolactone/analogs & derivatives , 4-Butyrolactone/genetics , 4-Butyrolactone/metabolism , Streptomyces/genetics , Bacterial Proteins/metabolism , Base Sequence , Binding Sites , DNA Footprinting , Gene Expression Regulation, Bacterial , Genes, Regulator , Models, Biological , Molecular Sequence Data , Receptors, Cell Surface/metabolism , Signal Transduction , Streptomyces/metabolism
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