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Northern blot and primer extension analyses revealed that theycnKJI operon and theycnL gene ofBacillus subtilis are transcribed from adjacent promoters that are divergently oriented. TheycnK andycnJ genes encode a DeoR-type transcriptional regulator and a membrane protein involved in copper uptake, respectively. DNA binding experiments showed that the YcnK protein specifically binds to theycnK -ycnL intergenic region, including a 16-bp direct repeat that is essential for the high binding affinity of YcnK, and that a copper-specific chelator significantly inhibits YcnK's DNA binding.lacZ reporter analysis showed that theycnK promoter is induced by copper limitation orycnK disruption. These results are consistent with YcnK functioning as a copper-responsive repressor that derepressesycnKJI expression under copper limitation. On the other hand, theycnL promoter was hardly induced by copper limitation, butycnK disruption resulted in a slight induction of theycnL promoter, suggesting that YcnK also repressesycnL weakly. Moreover, while the CsoR protein did not bind to theycnK -ycnL intergenic region,lacZ reporter analysis demonstrated thatcsoR disruption induces theycnK promoter only in the presence of intactycnK andcopZA genes. Since thecopZA operon is involved in copper export and repressed by CsoR, it appears that the constitutivecopZA expression brought bycsoR disruption causes intracellular copper depletion, which releases the repression of theycnKJI operon by YcnK.

journal of bacteriology

Direct and Indirect Regulation of the <i>ycnKJI</i> Operon Involved in Copper Uptake through Two Transcriptional Repressors, YcnK and CsoR, in Bacillus subtilis

Direct and Indirect Regulation of the ycnKJI Operon Involved in Copper Uptake through Two Transcriptional Repressors, YcnK and CsoR, in Bacillus subtilis