title: The human elongation factor 1 A-2 gene (EEF1A2): complete sequence and characterization of gene structure and promoter activity.
abstract: The eukaryotic elongation factor 1 A (eEF1A, formerly EF1alpha) is a key factor 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 report the isolation and sequencing of the gene (HGMW-approved symbol EEF1A2) coding for the human eEF1A-2 isoform. Furthermore, we characterize the gene structure and the activity of the promoter. Isolation of overlapping clones from human libraries revealed that the human eEF1A-2 gene spans approximately 10 kb and consists of eight exons. The intron-exon boundaries of human 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 upstream of the AUG codon. The start site maps to an adenine located within a consensus initiator element. Sequencing of a 2-kb 5'-flanking promoter region revealed no TATA element. However, several putative cis-regulatory elements were discovered. The 5'-promoter activity was characterized by transient transfection experiments. Progressive deletions of the upstream promoter region defined a minimal promoter region, ranging from -16 to +92, that is sufficient to drive transcription.
authors.affiliation: IMSB, University of Aarhus, Gustav Wieds vej 10 C, Arhus C, 8000, Denmark.
authors.forename: C
authors.initials: C
authors.lastname: Bischoff
authors.forename: S
authors.initials: S
authors.lastname: Kahns
authors.forename: A
authors.initials: A
authors.lastname: Lund
authors.forename: H F
authors.initials: HF
authors.lastname: Jørgensen
authors.forename: M
authors.initials: M
authors.lastname: Praestegaard
authors.forename: B F
authors.initials: BF
authors.lastname: Clark
authors.forename: H
authors.initials: H
authors.lastname: Leffers
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D005796:Genes / Q000235:genetics*; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D020648:Peptide Elongation Factor 1 / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020033:Protein Isoforms / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion; D014158:Transcription, Genetic
chemical_list: D020648:Peptide Elongation Factor 1; D020033:Protein Isoforms; D011993:Recombinant Fusion Proteins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase