Molecular analyses of metallothionein gene regulation

Metallothionein (MT) genes encode small proteins that chelate metal ions through metal-thiolate bonds with cysteine residues. MTs may have a role in cellular zinc homeostasis and metal detoxification. Congruent with these putative functions, MT gene transcription is induced by metals via multiple metal-responsive elements (MREs) present in the MT gene 5'-regulatory regions. This chapter mainly is focused on studies of the functional and physical interactions of MRE binding proteins with MT promoters from human and rainbow trout. In addition to mediating zinc induction, MREs may make important contributions to nonmetal induced promoter activity. In part, differential basal activity of MREs appears to be determined by sequence and position in the promoter. During zinc induction, increased functional MRE activity correlates with increased activity of mammalian MRE binding proteins by zinc treatment in vivo or in vitro, as detected by electrophoretic mobility shift assays. Interestingly, the addition of cadmium in vitro or in vivo has no detectable effect even though it strongly induces MT gene expression in the same time course. This raises questions about how the effects of cadmium are mediated by MREs. The molecular masses and MRE complex migration of the zinc-responsive factors we detect are consistent with mouse and human metal-responsive transcription factor (MTF) and expression of the MTF cDNAs increases co-transfected MT promoter activity in both mammalian and trout cell lines underlining the conservation of MRE binding factor function among diverse species.