FIG 3.

ZnuA is produced during, and required for, growth in low Zn conditions. (A) ΔznuA strains harboring pWH1266 alone or pWH1266 expressing znuA under the control of its native promoter were grown in either LB or LB + 30 μM TPEN, and 10 μg of cell lysate was added to each lane for immunoblotting. Equal loading of lanes is shown in Fig. S2 in the supplemental material, and the blot shown is representative of four independent experiments. (B) WT cells harboring pWH1266 and ΔznuA strains harboring an empty vector, a vector containing a WT copy of znuA, or a vector containing znuA with a substitution of A for H at residue 41 were grown in 40 μM TPEN and 75 μg/ml carbenicillin, and OD₆₀₀ was monitored over time. Expression of the ZnuA(H41A)-myc protein is demonstrated with an α-myc immunoblot in Fig. S2 in the supplemental material. Data are shown in biological triplicate ± SD and are representative of three independent experiments.