Figure 1.

eIF5A depletion yields a marked pause at the stop codon. (A) Growth of wild-type BY4741 and eIF5A temperature-sensitive mutant cells tif51A-1 and tif51A-3 in YPD media at permissive (25°C) and restrictive (37°C) temperatures. (B) Western blot analysis of eIF5A depletion. Cells were grown in YPD until the exponential phase and were then transferred to the indicated temperatures. Proteins were extracted and analysed by Western blot. The eIF5A protein was visualized with a specific anti-eIF5A antibody. (C) Outline of the 5PSeq method. (D) The metagene analysis displaying the abundance 5΄P intermediates in reads per million (rpm) in relation to the ORF start (left panel) and stop (right panel) codons.