The genomic DNA encoding the exons for the human neural phosphoprotein B-50 (GAP-43) was isolated using rat-based cDNA probes and oligonucleotides. Exons 2 and 3 were isolated from a genomic library, exon 1 was amplified by PCR on total genomic DNA. The gene consists of 3 exons and 2 large introns. The first exon encodes the N-terminal 10 amino acids of B-50 involved in membrane association of the protein. Exon 2 encodes the main part of the protein with the sites for protein kinase C-mediated phosphorylation and calmodulin binding, and includes a 10 amino acid residue insert not found in rodents. Exon 3 encodes the last 29 amino acid residues. The reported sequence extends the known cDNA structure to both the 5′ and 3′ ends. The 358 bp region upstream of the translational initiation codon, containing the main transcription starts, is purine-rich and does not include TATA or GC boxes. At the 3′ end potential polyadenylation signals were found 510 bp and 584 bp downstream of the stopcodon in exon 3. The 5′ end of the mRNA is heterogeneous in length, with primer extension products corresponding to a 5′ untranslated region of 159 and 343 bases. Northern hybridizations, however, indicate that the majority of B-50 mRNA has a shorter 5′ untranslated region, as was reported for the rat (Schrama et al., Soc. Neurosci. Abstr., 18 (1992) 333.4). The structural organization of the human gene is similar to that described for the rat (Grabczyk et al., Eur. J. Neurosci. 2 (1990) 822-827), and both translated and untranslated regions show a high degree of sequence homology to the rat gene.
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Acknowledgements. We thank Dr. J. H6ppener for helpful discussions, E.B.J. Ueffing for analysis of exon 3, A.J. van Rozen and N. Fresen for figures, and G. Peek for photography. H.B.N. is supported by the 'Prinses Beatrix Fonds'.
Copyright 2014 Elsevier B.V., All rights reserved.
- Gene structure
- Growth-associated protein
- Human gene
- Transcription start