Structural organization of the human alpha-galactosidase A gene: further evidence for the absence of a 3' untranslated region.

Abstract

Human a-galactosidase A (a-D-galactoside galactohydrolase; EC 3.2.1.22) is a lysosomal hydrolase encoded by a gene localized to the chromosomal region Xq22. The deficient activity of this enzyme results in Fabry disease, an X chromosome-hnked recessive disorder that leads to premature death in affected males. For studies of the structure and function of a-galactosidase A and for characterization of the genetic lesions in families with Fabry disease, the full-length cDNA was isolated, sequenced, and used to screen human genomic libraries. The 1393-base-pair full-length cDNA had a 60-nucleotide 5' untranslated region and encoded a precursor peptide of 429 amino acids including a signal peptide of 31 residues. Three overlapping A clones spanning 32 kilobases were identified that contained the entire ,12-kilobase chromosomal gene as well as -9 and -11 kilobases of 5' and 3' flanking sequence, respectively. The gene had seven exons. The genomic exonic and full-length cDNA sequences were identical. All intron-exon splice junctions conformed to the GT/AT consensus sequence. The 5' flanking region of this lysosomal housekeeping gene contained Spl and CCAAT box promoter elements as well as sequences corresponding to the activator protein 1 (AP1), octanucleotide ("OCTA"), and "core" enhancer elements. There was an upstream "HTF" island (Hpa I tiny fragments) followed by four direct repeats of the "chorion box" enhancer. The unique lack of a 3' untranslated sequence in the a-galactosidase A cDNA was confirmed by sequencing additional cDNA clones and the genomic 3' region.Human a-galactosidase A (a-D-galactoside galactohydrolase; EC 3.2

A 3′ splice site consensus sequence mutation in the intron 3 of the α-galactosidase a gene in a patient with Fabry disease

“…There was no difference in splicing donor site between patient and control (data not shown). The sequence result of at least 100 bases of the 5' end of intron 3 matched to the previous report (Bishop et al, 1988;Kornreich et al, 1989).…”

“…Recently, the full-length cDNA encoding human a-GalA has been isolated and characterized (Calhoun et al, 1985;Bishop et al, 1986Bishop et al, , 1988Kornreich et al, 1989). Since then molecular analysis of this disease has been reported.…”

Three α‐Galactosidase Genes of Trichoderma reesei Cloned by Expression in Yeast

“…The capability of some a-galactosidases to release a-galactosyl side groups from galactomannans has also been studied. Enzymes that can act on intact polymeric galactomannan have been isolated from seeds of Cyumopsis tetragonoloba (guar; Bulpin et al, 1990), Phaseolus vulgaris (French bean; Dhar et al, 1994) and Vigna rudiuta (mung bean; Dey, 1984) and from filamentous fungi such as Aspergillus tamarii (Civas et al, 1984b), Aspergillus niger (Adya and Elbein, 1977;Kaneko et al, 1991) and Trichoderma reesei (Zeilinger et al, 1993). Some cx-galactosidases have no activity on galactomannan such as the one described for Mortierella vinacea (Kaneko et al, 1990).…”

“…Genes encoding a-galactosidases have been isolated from several sources, such as from human (Bishop et al, 1988), plant seeds (Overbeeke eta!., 1989; Zhu and Goldstein, 19941, bacte-ria (Liljestrom and Liljestrom, 1987;Koyama et al, 1990;Russell et al, 1992;Aduse-Opoku et al, 1991), yeast (SumnerSmith et al, 1985) and filamentous fungi (den Herder et al, 1992;Shibuya et al, 1995). Based on similarities in primary structure and hydrophobic cluster analyses they have been grouped into three well conserved families in the general classification of glycosyl hydrolases (Henrissat and Bairoch, 1993).…”

Fabry Disease: Treatment and diagnosis

“…For this reason, genetic testing is carried out by direct sequencing of the gene coding for GLA (74). GLA gene is formed by seven exons, and the 1,393 bp full-length cDNA codes for a precursor peptide of 429 amino acids including the 31 residues' signal peptide (75). Alterations in the gene includes large rearrangements, splicing defects, insertions/deletions, and point mutations.…”