GENOMIC ORGANIZATION AND ISOFORMS OF THE MOUSE ELP GENE

Analysis was made on the genomic structure, functions, and expression of the mouse ELF gene, which codes for the embryonal long terminal repeat binding protein, Extensive screening of the cDNA library of embryonal carcinoma cells (EC cells) identified four isoforms of ELF: ELP1 (the original ELF isolate), ELP2, ELP3, and Ad4BP/SF1. Analysis of the genomic sequences revealed that these ELF isoforms were generated by alternative promoter usage and differential splicing. The mRNAs of isoforms initiated at four transcription start sites distributed on three exons, Sequence analysis of the four isoforms identified three polypeptides. The N-terminal portion of ELP1 and ELP2 was longer than ELP3, and Ad4BP/SF1 by 77 aa. The DNA-binding domain and region II were shared by all four isoforms. The C-terminal portion shared by ELP2, ELP3, and Ad4BP/SF1 was 131 aa in length, and that specific to ELP1 was 57 aa in length. The ELP3 and Ad4BP/SF1 isoforms were identical for the coding sequence, but the two differed at the 5' noncoding region, Region II and III domains of nuclear receptors mere thought to be involved in ligand-binding and transcriptional activation. ELP1, which lacked region III, functioned as a repressor, The isoforms carrying intact region II and region III functioned as transactivators. Expression of the four isoforms was studied in mouse tissues and in tissue culture cells by quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Complex patterns of expression of these isoforms were observed in various tissues. All four ELF isoforms were expressed only in EC cells.