YGR271C-A and YGR272C constitute a contiguous ORF required for growth at 37°C, growth on HU-containing media and cell cycle progression. (A) The ygr271c-aΔ, ygr272cΔ and ygr271c-aΔ/ygr272cΔ strains exhibit slow growth, Ts, and HU-sensitive phenotypes. Growth assays were carried out as in Figures 3 and 4. (B) YGR271C-A and YGR272C exhibit similarity to the A. gossypii ORF PABR143C (Brachat et al. 2003). By sequencing the genomic locus in S. cerevisiae, we determined that YGR271C-A and YGR272C constitute a single larger ORF. (C) Cell cycle analysis was done by arresting wild-type (BY4741) and ygr271c-aΔ/ygr272cΔ strains with α-factor (0 min) and then releasing them into pheromone-free media. Samples were analyzed for DNA content at 20-min intervals as indicated by flow cytometry. The 1C and 2C peaks denote cells with a 1N and 2N DNA content corresponding to cells in G1 or G2/M phase of the cell cycle, respectively.
