The dimeric repressor SoxR binds cooperatively to the promoter(s) regulating expression of the sulfur oxidation (sox) operon of Pseudaminobacter salicylatoxidans KCT001.
Sulfur oxidation in Pseudaminobacter salicylatoxidans KCT001 is rendered by the combined action of several enzymes encoded by a thiosulfate-inducible sox operon. In this study it has been conclusively demonstrated by insertional mutagenesis that the regulatory gene of this operon is soxR, which encodes a DNA-binding protein belonging to the ArsR-SmtB family. SoxR was found to bind to two promoter-operator segments within the sox cluster, of which the one (wx) located between soxW and soxX controls the expression of sulfur-oxidation genes soxX through soxD while the other, a bi-directional element (sv) located between soxS and soxV, controls the expression of soxVW in one direction and the putative regulatory cluster soxSRT in the other. In the case of the wx promoter the repressor was found to bind in a cooperative manner to two distinct binding sites having different affinities, while in the case of the sv promoter binding occurred at a symmetric dimeric site and involved a higher degree of cooperativity. The high degree of cooperativity observed in the binding of SoxR to its target sites seemed to be due to the propensity of SoxR monomers to form dimers. The apparent dissociation constants of the SoxR-operator complexes were in the nanomolar range, indicating relatively strong interactions. It was demonstrated using a reporter system in Escherichia coli that this high-affinity binding of SoxR led to efficient repression in trans. Thus the role of SoxR as a repressor of the sox operon has not only been conclusively established but it has also been shown that this repression is brought about through cooperative interactions of SoxR with dimeric binding sites that occlude the operon promoters.
Mandal
Sukhendu
S
Department of Microbiology, Bose Institute, P-1/12, CIT Scheme VII-M, Kolkata-700 054, India.
Chatterjee
Sujoy
S
Dam
Bomba
B
Roy
Pradosh
P
Das Gupta
Sujoy K
SK
D020561:Alphaproteobacteria / Q000235:genetics* / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics; D019281:Dimerization; D008969:Molecular Sequence Data; D009876:Operon / Q000235:genetics*; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic / Q000502:physiology; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013455:Sulfur / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; D015534:Trans-Activators; D014157:Transcription Factors; C070724:SoxR protein, Bacteria; D013455:Sulfur