Analysis of regulatory elements flanking metallothionein genes in Cd-tolerant fish (pike and stone loach).

From genomic libraries constructed for both pike and stone loach, clones were isolated containing the metallothionein genes from these two species of cadmium-tolerant fish. A single copy metallothionein gene was identified in each species by Southern blot analysis. Sequencing revealed that each gene consisted of three exons followed by polyadenylation signals at the 3' end. In the 5' flanking region, putative metal responsive elements were identified both close to the transcription start site and clustered distally approx. 500 bp upstream. Neither gene locus showed any homology with the glucocorticoid or interferon responsive elements that have been identified in mammalian species. The significance of the absence of such responsive elements and their replacement by additional metal responsive elements in the same location of the 5'-flanking region of the MT genes is discussed in relation to the organisation of the MT gene loci in (the Cd-sensitive) rainbow trout and higher mammalian species.