FIGURE 5.

Mapping a minimal zinc-responsive domain. A, schematic diagram of pL-Loz1GFP truncations. Shown are a C-terminal GFP tag, the double zinc finger domain (numbered black boxes), regions conserved in Loz1 homologs from other Schizosaccharomyces sp. (striped boxes), and introduced nuclear localization signal. The amino acids included in each truncation are also indicated. B and C, loz1Δ cells expressing the constructs shown in A were grown in ZL-EMM with or without a 200 μm zinc supplement. Cells were harvested, and crude protein extracts were prepared for immunoblot analysis (B), or total RNA was extracted for RNA blot analysis (C). D, loz1Δ cells co-expressing the constructs shown in A and reporter gene pTN-zrt1-lacZ (zrt1-lacZ) or empty vector pTN-lacZ (vector) were grown as described in B before β-galactosidase activity was measured by standard procedures. Error bars, S.E.