The Zinc-responsive Regulator Zur and Its Control of the znu Gene Cluster Encoding the ZnuABC Zinc Uptake System in Escherichia coli *

The synthesis of the Escherichia coli zinc transporter, encoded by the znuACB gene cluster, is regulated in response to the intracellular zinc concentration by the zur gene product. Inactivation of the zur gene demonstrated that Zur acts as a repressor when binding Zn 2+. Eight chromosomal mutant zur alleles were sequenced to correlate the loss of Zur function with individual mutations. Wild-type Zur and ZurΔ46–91 formed homo- and heterodimers. Dimerization was independent of metal ions since it also occurred in the presence of metal chelators. Using an in vivo titration assay, the znu operator was narrowed down to a 31-base pair region overlapping the translational start site of znuA. This location was confirmed by footprinting assays. Zur directly binds to a single region comprising a nearly perfect palindrome. Zinc chelators completely inhibited and Zn 2+ in low concentrations enhanced DNA binding of Zur. No evidence for autoregulation of Zur was found. Zur binds at least 2 zinc ions/dimer specifically. Although most of the mutant Zur proteins bound to the znu operator in vitro, no protection was observed in in vivo footprinting experiments. Analysis of the mutant Zur proteins suggested an amino-terminal DNA contact domain around residue 65 and a dimerization and Zn 2+ -binding domain toward the carboxyl-terminal end.