Puf3-dependent protein expression is disrupted in Mkt1 and Pbp1 truncation strains. (A) Schematic representation of Flag-tagged Pbp1 truncation strains assayed for their capacity to express mitochondrial proteins and grow in YPL medium. (B) Western blot depicting Cox2, Pbp1-Flag, and Por1 protein levels during growth in YPD and following switch to YPL medium. Pbp1Δ491-570 and Pbp1Δ491-513 cells had diminished Cox2 and Pbp1-Flag protein levels compared with WT. (C) Growth curves of indicated strains collected using an automatic plate reader during incubation in YPL medium at 30°C. OD₆₀₀ measurements were obtained every 30 min. Note: Pbp1Δ491-570 and Pbp1Δ491-513 cells had reduced growth. Representative traces from a single experiment are depicted (n = 3 per group). (D) Western blot depicting Cox2, Pbp1-Flag, and Por1 protein levels during growth in YPD and following switch to YPL medium. Pbp1Δ500-514 cells had decreased Cox2 and Pbp1-Flag protein levels compared with WT. (E) Growth curves of indicated strains during incubation in YPL medium obtained using the method described in C. Note: Pbp1Δ500-514 cells had reduced growth. Representative traces from a single experiment are depicted (n = 3 per group). (F) Western blot depicting Cox2, Mkt1-Flag, and Por1 protein levels during growth in YPD and following switch to YPL medium. Mkt1 truncation strains had reduced Cox2 protein abundance compared with WT. (G) Growth curves of indicated strains during incubation in YPL medium using method described in C. Note: Mkt1 truncation strains had reduced growth. Representative traces from a single experiment are depicted (n = 3 per group). Source data are available for this figure: SourceData FS3.