The ManR specifically binds to the promoter of a Nramp transporter gene in Anabaena sp. PCC 7120: a novel regulatory DNA motif in cyanobacteria.
The Anabaena sp. PCC 7120 ManR and a homologous protein of MntH were identified by BLAST search. Recombinant ManR protein was overexpressed in Escherichia coli and purified by an immobilized metal (Ni) affinity chromatography. Electrophoretic mobility shift assays revealed that ManR specifically bound to the promoter region of the mntH gene. Site-directed mutagenesis experiments demonstrated that the specific recognition site for ManR is TATGAAAAGAATATGAGAA, which is composed of two direct repeats of the consensus sequence (T/A)ATGA(G/A)A(A/G). This is a novel regulatory DNA motif in cyanobacteria, indicating that the expression of mntH was regulated by a two-component Mn(2+)-Sensing System containing ManR in Anabaena sp. PCC 7120. To date, this specific pathway of regulating mntH expression has only been found in cyanobacteria.
Huang
Wei
W
Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing 100084, PR China.
Wu
Qingyu
Q
D000595:Amino Acid Sequence; D017033:Anabaena / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D027682:Cation Transport Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016384:Consensus Sequence; D000458:Cyanobacteria / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D000099173:Natural Resistance-associated Macrophage Protein 1
D001426:Bacterial Proteins; D002352:Carrier Proteins; D027682:Cation Transport Proteins; D029968:Escherichia coli Proteins; D000099173:Natural Resistance-associated Macrophage Protein 1; C492412:MntH protein, E coli; C433793:MntH protein, bacteria