Structural and functional analysis of the rat metallothionein III genomic locus

Metallothionein III (MT III) has been reported to suppress neuronal growth in a rat in vitro model system. The protein and its specific mRNA are detected predominantly in the brain, differentiating MT III from the well-characterised archetypal metallothioneins. Isolation, sequencing and functional analysis of the rat MT III genomic locus indicated that, although the organisation of the gene was conserved between MT III and the more conventional metallothioneins, the 5' flanking region of the MT III gene was distinct. Within this region, a number of putative regulatory elements were identified, including the metal regulatory elements (MREs) characteristic of metallothionein promoters. However, despite their conservation in sequence with active elements, the MREs of MT III were unresponsive to zinc. A 'silencing element' was revealed within a 250 bp section of the MT III promoter which suppressed gene expression in two brain cell lines. The operation of this silencing region in conjunction with the inactive MREs may explain the distinct expression profile observed for MT III within the central nervous system and during neuronal development.