Metal specificity of cyanobacterial nickel-responsive repressor InrS: cells maintain zinc and copper below the detection threshold for InrS.

ABSTRACT

InrS is a Ni(II)-responsive, CsoR/RcnR-like, DNA-binding transcriptional repressor of the nrsD gene, but the Ni(II) co-ordination sphere of InrS is unlike Ni(II)-RcnR. We show that copper and Zn(II) also bind tightly to InrS and in vitro these ions also impair InrS binding to the nrsD operator-promoter. InrS does not respond to Zn(II) (or copper) in vivo after 48 h, when Zn(II) sensor ZiaR responds, but InrS transiently responds (1 h) to both metals. InrS conserves only one (of two) second co-ordination shell residues of CsoR (Glu98 in InrS). The allosteric mechanism of InrS is distinct from Cu(I)-CsoR and conservation of deduced second shell residues better predicts metal specificity than do the metal ligands. The allosteric mechanism of InrS permits greater promiscuity in vitro than CsoR. The factors dictating metal-selectivity in vivo are that KNi(II) and ΔG(C)(Ni(II)-InrS·DNA) are sufficiently high, relative to other metal sensors, for InrS to detect Ni(II), while the equivalent parameters for copper may be insufficient for copper-sensing in Synechocystis (at 48 h). InrS K(Zn(II)) (5.6 × 10(-13) M) is comparable to the sensory sites of ZiaR (and Zur), but ΔG(C)(Zn(II)-InrS·DNA) is less than ΔG(C)(Zn(II)-ZiaR·DNA) implying that relative to other sensors, ΔG(C)(Zn(II)-Sensor·DNA) rather than K(Zn(II)) determines the final detection threshold for Zn(II).