Cloning of human AMP deaminase isoform E cDNAs. Evidence for a third AMPD gene exhibiting alternatively spliced 5'-exons.
Higher eukaryotes express multiple isoforms of AMP deaminase (EC 3.5.4.6). In humans, four AMP deaminase variants, termed M (muscle), L (liver), E1, and E2 (erythrocyte) can be distinguished by a variety of biochemical and immunological criteria. Previous molecular studies have reported two genes, AMPD1 and AMPD2, that produce isoform M and L transcripts, respectively. This study identifies a third human AMP deaminase gene, AMPD3. Nucleotide sequence alignments between AMPD3 cDNAs isolated from several human libraries indicate three different extreme 5'-ends. Alternate forms of the AMPD3 cDNAs contain a common 2301-bp open reading frame (ORF) and 3'-untranslated region of 1245 bp. Two of the three forms, however, exhibit additional 5'-end nucleotide sequences that would extend their respective ORFs by 21 and 27 nucleotides. RNase protection analyses and the partial characterization of human AMPD3 genomic clones demonstrate alternative splicing of three different 5'-terminal exons. Western blot analyses detect anti-E-specific immunoreactivity in affinity-purified extracts derived from the bacterial expression of a truncated AMPD3 cDNA. These results are discussed in relation to AMP deaminase isoform diversity.
Mahnke-Zizelman
D K
DK
Department of Cellular Biology and Anatomy, Medical College of Wisconsin, Milwaukee 53226.
Sabina
R L
RL
D000659:AMP Deaminase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D003001:Cloning, Molecular / Q000379:methods; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004273:DNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D004912:Erythrocytes / Q000201:enzymology; D005091:Exons; D014644:Genetic Variation; D015698:Genomic Library; D006801:Humans; D007438:Introns; D007527:Isoenzymes / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015459:Leukemia-Lymphoma, Adult T-Cell; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D009132:Muscles / Q000201:enzymology; D016366:Open Reading Frames; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D007527:Isoenzymes; D011994:Recombinant Proteins; D004247:DNA; D000659:AMP Deaminase