THE ENTIRE GENOMIC SEQUENCE AND CDNA EXPRESSION OF MOUSE ALPHA-GALACTOSIDASE-A

The full-length cDNA and genomic sequences encoding mouse alpha-galact osidase A (alpha Gal A; EC 3.2.1.22), a lysosomal galactohydrolase, we re isolated and characterized. The cDNA's open reading frame encoded 4 19 amino acids and had 82% nucleotide (nt) and 78% amino acid identity with the human sequence, although the carboxy terminus of the mouse a lpha-Gal A polypeptide was 10 amino acids shorter. The functional inte grity of the mouse cDNA was demonstrated by transient expression in CO S-1 cells. Northern analysis revealed two mRNA species of about 1.6 an d 3.4 kb due to alternative polyadenylation signals. The entire 14.4-k b mouse genomic sequence was determined; each of its seven exons was i nterrupted by intronic sequence at the identical positions as the exon s in the human gene. The mouse 5' flanking region (250 nt) had one Sp1 site, five CAAT boxes, and no TATA box and had 67% identity with the human promoter region. The gene contained 18 complete or partial Alu-r epetitive elements (13 type 1 and 5 type 2 repeats), and three putativ e functional AATAAA consensus polyadenylation signals were identified 72, 1668, and 1682 nt after the TAA termination codon. Use off the 72- nt site and the 1688 and/or 1682 sites were consistent with the shorte r and longer transcripts. The availability of the full-length cDNA and genomic sequence encoding mouse alpha-Gal A should facilitate structu re/function studies of this lysosomal glycosidase and the construction of alpha-Gal A-deficient mice by targeted gene disruption. (C) 1996 A cademic Press, Inc.