HUMAN GALACTOCEREBROSIDASE GENE - PROMOTER ANALYSIS OF THE 5'-FLANKING REGION AND STRUCTURAL ORGANIZATION

Galactocerebrosidase (GALC; EC 3.2.1.46) is a lysosomal enzyme which h ydrolyzes several galactolipids and the deficiency of GALC is responsi ble for Krabbe disease. Recently, we cloned cDNAs for human and murine GALC. In this study we characterized the genomic organization and the promoter of the human gene, The gene was about 60 kb in length and co nsisted of 17 exons as reported by Luzi et al. [1]. DNA sequence analy sis showed that the 5'-flanking region of the first exon was GC-rich a nd had not typical TATA-box but ten GC-box-like sequences within a 200 bp sequence upstream from the initiation codon. Another inframe ATG, which has better Kozak consensus sequence, was found at 48 bp upstream to the first ATG reported [1]. Promoter analysis using a luciferase a ssay in COS 7 cells showed that the -149 to -112 nucleotide (from the initiation codon A) region has dominant promoter activity. In this reg ion three GC-box-like sequence and one YY1 binding site were detected. Primer extension revealed several transcription start sites within th e region of -146 to -103 nucleotide. In this study we firstly demonstr ated that the YY1 binding site and subsequent GC-box-like sequences co uld be a promoter in a housekeeping gene. (C) 1998 Elsevier Science B. V.