Mucopolysaccharidosis IV A: Molecular cloning of the human N-Acetylgalactosamine-6-sulfatase gene (GALNS) and analysis of the 5[prime]-flanking region

Journal Article · Mon Feb 28 23:00:00 EST 1994 · Genomics; (United States)

DOI:https://doi.org/10.1006/geno.1994.1132· OSTI ID:6789611

Nakashima, Yoshihiro; Tomatsu, Shunji; Hori, Toshinori; Fukuda, Seiji; Sukegawa, Kazuko; Kondo, Naomi; Suzuki, Yasuyuki; Shimozawa, Nobuyuki; Orii, Tadao ^[1]

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1. Gifu Univ. School of Medicine (Japan)

Mucopolysaccharidosis IV A (MPS IV A) is the result of a genetic deficiency in a lysosomal hydrolase, N-acetylgalactosamine-6-sulfatase (GALNS). To investigate MPS IV A patients at the level of the genome, the authors analyzed the structure of the human GALNS-encoding gene. From the genomic library of a normal subject in [lambda]EMBL3, they isolated five overlapping clones covering the coding region of the GALNS cDNA and determined the structural organization. The gene is about 50 kb long and contains 14 exons. The 5[prime]-flanking region lacks a canonical TATA box and CCAAT sequences, but is G+C-rich (70.5%), with four GC boxes, characteristic of a housekeeping gene promoter. The transcription initiation site was determined by primer extension analysis, using RNA from human liver and Hela cells. Transcription was found to initiate at a few sites, the major ones being 58 and 22 bp upstream of the translation initiation codon. The 5[prime]-flanking region had promoter activity by transient expression, determined using a CAT assay. In addition, this region retained promoter activity, even in reverse orientation. The region -98 to -1 upstream of the ATG codon was defined by deletion analysis to be a minimal promoter. One GC box in this region is likely to be a binding site of a regulatory element. 36 refs., 4 figs., 1 tab.

• OSTI ID: 6789611
• Journal Information: Genomics; (United States), Journal Name: Genomics; (United States) Vol. 20:1; ISSN GNMCEP; ISSN 0888-7543
• Country of Publication: United States
• Language: English

Related Subjects

550400* -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMINES
CARBOHYDRATES
CLONING
DISEASES
DNA HYBRIDIZATION
DNA SEQUENCING
DNA-CLONING
ENZYMES
ESTERASES
GENE MUTATIONS
HYBRIDIZATION
HYDROLASES
METABOLIC DISEASES
MUCOPOLYSACCHARIDES
MUTATIONS
ORGANIC COMPOUNDS
POLYSACCHARIDES
PROTEINS
SACCHARIDES
STRUCTURAL CHEMICAL ANALYSIS