An iron-sulfur center essential for transcriptional activation by the redox-sensing SoxR protein.
The soxRS oxidative stress regulon of Escherichia coli is triggered by superoxide (O2.-) generating agents or by nitric oxide through two consecutive steps of gene activation. SoxR protein has been proposed as the redox sensing gene activator that triggers this cascade of gene expression. We have now characterized two forms of SoxR: Fe-SoxR contained non-heme iron (up to 1.6 atoms per monomer); apo-SoxR was devoid of Fe or other metals. The spectroscopic properties of Fe-SoxR indicated that it contains a redox active iron-sulfur (FeS) cluster that is oxidized upon extraction from E. coli. Fe-SoxR and apo-SoxR bound the in vivo target, the soxS promoter, with equal affinities and protected the same region from DNase I in vitro. However, only Fe-SoxR stimulated transcription initiation at soxS in vitro > 100-fold, similar to the activation of soxS expression in vivo. This stimulation occurred at a step after the binding of RNAP and indicates a conformational effect of oxidized Fe-SoxR on the soxS promoter. The variable redox state of the SoxR FeS cluster may thus be employed in vivo to modulate the transcriptional activity of this protein in response to specific types of oxidative stress.
Hidalgo
E
E
Department of Molecular and Cellular Toxicology, Harvard School of Public Health, Boston, MA 02115.
Demple
B
B
D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D004269:DNA, Bacterial; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D004926:Escherichia coli; D007501:Iron / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000737:chemistry / Q000378:metabolism*; D008667:Metalloproteins / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D019159:Nonheme Iron Proteins; D010084:Oxidation-Reduction; D011401:Promoter Regions, Genetic; D013057:Spectrum Analysis; D013455:Sulfur / Q000378:metabolism; D014157:Transcription Factors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D015533:Transcriptional Activation
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D007506:Iron-Sulfur Proteins; D008667:Metalloproteins; D019159:Nonheme Iron Proteins; D014157:Transcription Factors; C070724:SoxR protein, Bacteria; D013455:Sulfur; D007501:Iron; D012321:DNA-Directed RNA Polymerases