FIG. 2.

Nhp6 protein mediates transcription. (A) Haploid segregants from diploid strain NB40 (Δnhp6a::URA3/+ Δnhp6b::LEU2/+ his4-912δ/his4-912δ lys2-128δ/lys2-128δ) were grown on solid rich medium and replica plated to histidine-free medium to assess functional transcription from the his4-912δ reporter gene. The lower panels indicate NHP6A/B genotypes as indicated by Ura and Leu phenotypes. (B) Representative segregants from AW11-9a × NB6040 with genotypes suc2ΔUAS Δnhp6a/b harboring the NHP6A plasmid p314-NV5 or the pRS314 vector, SUC2 Δnhp6a/b, and suc2ΔUAS NHP6A NHP6B were grown on solid rich medium at 28°C, replica plated to rich medium containing glucose or sucrose (plus antimycin A at 1 μg/ml) as the carbon source, and incubated 3 days at 23°C. (C) Representative segregants from FY711 × NB39-18a with genotypes snf5-5::URA3 Δnhp6a/b with or without p314-NV5, SNF5 NHP6A NHP6B, and snf5-5 NHP6A NHP6B were grown on solid rich medium at 23°C, replica plated to rich medium, with or without 1 M sorbitol, containing glucose or sucrose (plus antimycin A) as the carbon source, and incubated for 6 days at 23 and 33°C. (D) Representative segregants from NB39-24b × (FY300 × NB39-11a segregant) were replica plated onto rich medium and incubated at the indicated temperatures. (E) Representative PPR2 segregants from NB2Δp-28b × NB40-1aF with genotypes Δnhp6a::URA3 Δnhp6b::LEU2, Δnhp6a::URA3 NHP6B, and Δnhp6a::URA3 Δnhp6b::LEU2 with POB3-NHP6A in place of POB3 were replica plated to uracil-free medium containing 6AU (100 μg/ml) and incubated at 28°C.