The human elongation factor 1 A-2 gene (EEF1A2): Complete sequence and characterization of gene structure and promoter activity

The eukaryotic elongation factor 1 A (eEF1A, formerly EF1 alpha) is a key f actor in protein synthesis, where it promotes the transfer of aminoacylated tRNAs to the A site of the ribosome, Two differentially expressed isoforms of eEF1A, designated eEF1A-1 and eEF1A-2, are found in mammals. Here we re port the isolation and sequencing of the gene (HG;MW-approved symbol EEF1A2 ) coding for the human eEF1A-2 isoform. Furthermore, we characterize the ge ne structure and the activity of the promoter. Isolation of overlapping clo nes from human libraries revealed that the human eEF1A-2 gene spans approxi mately 10 kb and consists of eight exons, The intron-exon boundaries of hum an EEF1A2 and EEF1A1 are conserved, yet the gene of the eEF1A-2 isoform is larger than the eEF1A-1 gene because of enlarged introns. Primer extension analysis identified the predominant transcription start site 166 bp upstrea m of the AUG codon, The start site maps to an adenine located within a cons ensus initiator element. Sequencing of a 2-kb 5'-flanking promoter region r evealed no TATA element, However, several putative cis-regulatory elements were discovered. The 5'-promoter activity was characterized by transient tr ansfection experiments. Progressive deletions of the upstream promoter regi on defined a minimal promoter region, ranging from -16 to +92, that is suff icient to drive transcription, (C) 2000 Academic Press.