Zn(2+)-responsive repressor ZiaR and Co(2+)-responsive activator CoaR modulate production of P(1)-type Zn(2+)- (ZiaA) and Co(2+)- (CoaT) ATPases respectively. What dictates metal selectivity? We show that Delta ziaDeltacoa double mutants had similar Zn(2+) resistance to Deltazia single mutants and similar Co(2+) resistance to Deltacoa single mutants. Controlling either ziaA or coaT with opposing regulators restored no resistance to metals sensed by the regulators, but coincident replacement of the deduced cytosolic amino-terminal domain CoaT(N) with ZiaA(N) (in ziaR-(p) ziaA-ziaA(N)coaT) conferred Zn(2+) resistance to DeltaziaDeltacoa, Zn(2+) content was lowered and residual Co(2+) resistance lost. Metal-dependent molar absorptivity under anaerobic conditions revealed that purified ZiaA(N) binds Co(2+) in a pseudotetrahedral two-thiol site, and Co(2+) was displaced by Zn(2+). Thus, the amino-terminal domain of ZiaA inverts the metals exported by zinc-regulated CoaT from Co(2+) to Zn(2+), and this correlates simplistically with metal-binding preferences; K(ZiaAN) Zn(2+) tighter than Co(2+). However, Zn(2+) did not bleach Cu(+)-ZiaA(N), and only Cu(+) co-migrated with ZiaA(N) after competitive binding versus Zn(2+). Bacterial two-hybrid assays that detected interaction between the Cu(+)-metallochaperone Atx1 and the amino-terminal domain of Cu(+)-transporter PacS(N) detected no interaction with the analogous, deduced, ferredoxin-fold subdomain of ZiaA(N). Provided that there is no freely exchangeable cytosolic Cu(+), restricted contact with the Cu(+)-metallochaperone can impose a barrier impairing the formation of otherwise favoured Cu(+)-ZiaA(N) complexes.