Fig. 1
From: Temperature-dependent regulation of upstream open reading frame translation in S. cerevisiae
Changes in temperature result in changes in relative expression of firefly luciferase (F-Luc) reporters with AUG and near cognate codons (NCCs) as start sites. a Schematic of the reporters used in the study. P, promoter; T, terminator. Renilla luciferase (R-Luc) mRNA was produced from the ADH promoter and HIS terminator, and Firefly luciferase (F-Luc) mRNA was produced from the GPD promoter and CYC terminator. “XXX” in the F-Luc test reporter represents a start codon that varies from AUG by 1 bp (NCC). The relative expression from Firefly luciferase with respect to Renilla luciferase was calculated for both test (F-LucXXX/R-LucAUG) and control (F-LucAUG/R-LucAUG) reporters. The normalized expression was then calculated as the ratio of relative F-LucXXX expression from the test reporter to relative F-LucAUG expression from the control reporter (XXX/AUG ratio). b Normalized F-LucUUG expression (UUG/AUG ratio) was measured in yeast strain BY4741 at different temperatures. c Normalized expression from F-Luc reporters (XXX/AUG ratio) with different NCCs as start sites in BY4741 cells cultured at multiple temperatures. d Same as c, except the experiments were done in Δupf1 cells. For b–d, the results are represented as averages of at least four biological replicates with error bars representing standard deviation of mean. Each circle on the individual columns represents one biological replicate. For b-d, the asterisk denotes p values < 0.05 calculated by Student’s t test when compared with 30 °C