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. 2016 Jun 2;44(16):7722–7741. doi: 10.1093/nar/gkw506

Figure 1.

Figure 1.

Identification of HOP1 in a genetic screen for high-copy suppressors of the zip1 pch2 checkpoint defect using a pch2-lacZ construct as a reporter for meiotic prophase arrest. (A) Schematic representation of a centromeric plasmid (pSS51) carrying the PCH2 promoter and the coding sequence for the first N-terminal 90 amino acids fused in frame with the bacterial lacZ gene. (B) Dityrosine fluorescence and β-galactosidase assays of the indicated strains transformed with the pSS51 plasmid after 48 h on sporulation plates. (C) Kinetics of meiotic divisions and β-galactosidase activity in meiotic time courses of wild-type and zip1 strains containing the pSS51 plasmid. (D) Schematic representation of the substitution of the PCH2 gene for the pch2-lacZ construct at the genomic locus. (E) Dityrosine fluorescence and β-galactosidase assays of the indicated strains after 48 h on sporulation plates. (F) Scheme of the genetic screen. (G) Representative β-galactosidase assay of a plate from zip1 pch2Δ-lacZ transformed with the genomic high-copy library. Black triangles indicate positive controls deliberately placed at known positions on every plate of the screen. The red arrow points to a positive ‘blue’ candidate. (H) Dityrosine fluorescence and β-galactosidase assays of the zip1 pch2Δ-lacZ strain transformed with the indicated plasmids. Strains for (B) and (C) are: BR2495 (wild type), MY63 (zip1), DP174 (zip1 dot1) and S3483 (ndt80). Strains for (E), (F), (G) and (H) are: DP221 (zip1 pch2Δ-lacZ) and DP228 (zip1 pch2Δ-lacZ/PCH2). DP221 was transformed with empty vector or the indicated high-copy plasmids.