Functional analysis of the cDNA encoding human tyrosinase precursor

Abstract

The DNA segment harboring the promoter region and the exon 1 of the human tyrosinase gene has been cloned and characterized. Sequence analysis reveals the amino-terminal half of tyrosinase molecule including a signal peptide, of which six amino acid residues are not represented in the tyrosinase cDNA, pHT gamma 1 [Shibahara et al. (1988) Tohoku J. Exp. Med. 156, 403-414]. We therefore constructed the expression plasmid containing the human tyrosinase precursor cDNA, and introduced it into mouse amelanotic melanoma cells. Both tyrosine hydroxylase and dopa oxidase activities were expressed only in the cells transfected with such a full-length cDNA, providing direct evidence that tyrosinase actually possesses a dual catalytic activity.