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PMID:3536930

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Citation

Ecker, DJ, Butt, TR, Sternberg, EJ, Neeper, MP, Debouck, C, Gorman, JA and Crooke, ST (1986) Yeast metallothionein function in metal ion detoxification. J. Biol. Chem. 261:16895-900

Abstract

A genetic approach was taken to test the function of yeast metallothionein in metal ion detoxification. A yeast strain was constructed in which the metallothionein locus was deleted (cup1 delta). The cup1 delta strain was complemented with normal or mutant metallothionein genes under normal or constitutive regulatory control on high copy episomal plasmids. Metal resistance of the cup1 delta strain with and without the metallothionein-expressing vectors was analyzed. The normally regulated metallothionein gene conferred resistance only to copper (1000-fold); constitutively expressed metallothionein conferred resistance to both copper (500-fold) and cadmium (1000-fold), but not to mercury, zinc, silver, cobalt, nickel, gold, platinum, lanthanum, uranium, or tin. Two mutant versions of the metallothionein gene were constructed and tested for their ability to confer metal resistance in the cup1 delta background. The first had a deletion of a highly conserved amino acid sequence (Lys-Lys-Ser-Cys-Cys-Ser). The second was a hybrid gene consisting of the sequences coding for the first 20 amino acids of the yeast protein fused to the monkey metallothionein gene. Expression of these genes under the CUP1 promoter provided significant protection from copper, but none of the other metals tested. These results demonstrate that there is significant flexibility in the structural requirements for metallothionein to function in copper detoxification and that yeast metallothionein is also capable of detoxifying cadmium under conditions of constitutive expression.

Links

PubMed

Keywords

Amino Acid Sequence; Base Sequence; Genes; Genes, Fungal; Metallothionein/genetics; Metallothionein/metabolism; Metals/pharmacology; Mutation; Plasmids; Saccharomyces cerevisiae/drug effects; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae/metabolism

Significance

Annotations

Gene product Qualifier GO Term Evidence Code with/from Aspect Extension Notes Status

YEAST:MTCU2

involved_in

GO:0010273: detoxification of copper ion

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:MTCU2

involved_in

GO:0071585: detoxification of cadmium ion

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:MTCU1

involved_in

GO:0010273: detoxification of copper ion

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete

YEAST:MTCU1

involved_in

GO:0071585: detoxification of cadmium ion

ECO:0000315: mutant phenotype evidence used in manual assertion

P

Seeded From UniProt

complete


See also

References

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