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PMID: 10950927 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

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

Genomics ·Vol. 68 ·No. 1 ·2000-08-15 ·Pages 63-70

Bischoff C, Kahns S, Lund A, Jørgensen HF, Praestegaard M, Clark BF, Leffers H

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.

MeSH Terms
Base Sequence Cell Line Chloramphenicol O-Acetyltransferase/genetics,metabolism DNA/chemistry,genetics,isolation & purification Exons Genes/genetics Humans Introns Molecular Sequence Data Peptide Elongation Factor 1/genetics Promoter Regions, Genetic/genetics Protein Isoforms/genetics Recombinant Fusion Proteins/genetics,metabolism Sequence Analysis, DNA Sequence Deletion Transcription, Genetic
Chemicals
Peptide Elongation Factor 1 Protein Isoforms Recombinant Fusion Proteins DNA Chloramphenicol O-Acetyltransferase
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Bischoff C
IMSB, University of Aarhus, Gustav Wieds vej 10 C, Arhus C, 8000, Denmark.
Kahns S
Lund A
Jørgensen H F
Praestegaard M
Clark B F
Leffers H
Article Info
Journal
Genomics
Abbr.
Genomics
ISSN
0888-7543
Published
2000-08-15
Pages
63-70
Language
English
Region
United States
NLM ID
8800135
Subset
IM
Databases
GENBANK
AF163763
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