Differential transcription of the two Saccharomyces cerevisiae genes encoding elongation factor 2

Abstract

The single polypeptide chain of elongation factor 2 (EF-2) is encoded by two Saccharomyces cerevisiae genes ( EFT1 and EFT2) with unique flanking sequences. One gene is necessary and either is sufficient for cell viability. In the present work, we have analyzed the transcription of EFT1 and EFT2. Although both genes harbor multiple transcription start points, EFT1 initiates primarily at residue C at −39 and EFT2 at residue C at −37. Several candidate TATA boxes were identified in each gene. Deletion analysis employing lacZ promoter fusions demonstrated that the promoter for EFT2 is located within a 79-bp region beginning 335 nucleotides (nt) upstream from the start ATG codon. This region contains two overlapping sequences with homology to the consensus binding site for the yeast transcription factor, Rap1p/Grf 1p/Tuf. In contrast, the sequences essential for the transcription of EFT1 were localized to the region between the start ATG and the stop codon of the VPS17 gene that terminates 267 nt upstream on the same strand. Analysis of promoter strengths using lacZ fusions indicated that the promoter for EFT2 is approx. 2.5-fold more active than that of EFT1. Analysis of the steady-state levels of mRNAs revealed that EFT2 contributes approx. 70% of the total EF-2 mRNA while the remaining 30% is produced by EFT1 We conclude that the difference in expression of EFT1 and EFT2 is due to the differential transcription of their promoters.