Abstract
Addition of copper to yeast cells leads to the induction of a low molecular weight, cysteine-rich protein that binds copper. This protein, termed copper chelatin or thionein, is related to the metallothionein family of proteins that are induced in response to cadmium and zinc in vertebrate cells. We have determined the structure of the yeast copper-binding protein by DNA sequence analysis of the gene. Although the 6573-dalton yeast protein is substantially divergent from vertebrate metallothioneins, the arrangement of 12 cysteine residues, which is a hallmark of metal-binding proteins, is partially conserved. We analyzed the regulatory DNA sequence of the gene by fusing it with the Escherichia coli galactokinase gene and assaying the levels of enzyme activity in yeast in response to copper. The transcriptional activation has a specific requirement for copper. Zinc, cadmium, and gold were unable to regulate the galactokinase activity. The yeast copper metallothionein regulatory sequences represent a previously unreported class of yeast promoter that is regulated by copper.
MeSH Terms
Amino Acid Sequence
Base Sequence
Copper
DNA, Fungal/genetics
Gene Expression Regulation
Genes
Metallothionein/genetics
Operon
Saccharomyces cerevisiae/genetics
Transcription, Genetic
Chemicals
DNA, Fungal
Copper
Metallothionein
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Butt T R
Sternberg E J
Gorman J A
Clark P
Hamer D
Rosenberg M
Crooke S T
References (23)
23 references, click to expand
-
Copper-induced synthesis of copper-chelatin in rat liver.
Arch Biochem Biophys. 1975 Sep;170(1):267-77
PMID: 1164032
-
Cloning and expression of a yeast copper metallothionein gene.
Gene. 1984 Jan;27(1):23-33
PMID: 6370791
-
Homologous copper(I)-(thiolate)2-chromophores in yeast copper thionein.
Biochim Biophys Acta. 1977 Aug 23;493(2):465-77
PMID: 329891
-
Prediction of the secondary structure of proteins from their amino acid sequence.
Adv Enzymol Relat Areas Mol Biol. 1978;47:45-148
PMID: 364941
-
Thionein gene expression in Cd++-variants of the CHO cell: correlation of thionein synthesis rates with translatable mRNA levels during induction, deinduction, and superinduction.
Nucleic Acids Res. 1979 Sep 11;7(1):271-88
PMID: 493142
-
Sequencing end-labeled DNA with base-specific chemical cleavages.
Methods Enzymol. 1980;65(1):499-560
PMID: 6246368
-
Isolation of a yeast centromere and construction of functional small circular chromosomes.
Nature. 1980 Oct 9;287(5782):504-9
PMID: 6999364
-
Induction of metallothionein in HeLa cells by dexamethasone and zinc.
Eur J Biochem. 1981 Jan;113(2):267-72
PMID: 7202410
-
Structure of mouse metallothionein-I gene and its mRNA.
Nature. 1981 Jul 16;292(5820):267-9
PMID: 7254320
-
The genes for fifteen ribosomal proteins of Saccharomyces cerevisiae.
J Biol Chem. 1981 Oct 10;256(19):10176-83
PMID: 6268628
-
The actin gene in yeast Saccharomyces cerevisiae: 5' and 3' end mapping, flanking and putative regulatory sequences.
Nucleic Acids Res. 1981 Dec 11;9(23):6339-50
PMID: 6275358
-
Human metallothionein genes--primary structure of the metallothionein-II gene and a related processed gene.
Nature. 1982 Oct 28;299(5886):797-802
PMID: 7133118
-
Tandem gene amplification mediates copper resistance in yeast.
Proc Natl Acad Sci U S A. 1982 Sep;79(17):5342-6
PMID: 6291039
-
Dramatic growth of mice that develop from eggs microinjected with metallothionein-growth hormone fusion genes.
Nature. 1982 Dec 16;300(5893):611-5
PMID: 6958982
-
cDNA cloning and nucleotide sequence comparison of Chinese hamster metallothionein I and II mRNAs.
Nucleic Acids Res. 1983 Feb 11;11(3):901-10
PMID: 6687636
-
Regulation of a metallothionein-growth hormone hybrid gene in bovine papilloma virus.
Proc Natl Acad Sci U S A. 1983 Jan;80(2):397-401
PMID: 6300840
-
Model for mammalian metallothionein structure.
Proc Natl Acad Sci U S A. 1983 Mar;80(6):1501-5
PMID: 6572910
-
The new yeast genetics.
Nature. 1983 Sep 29-Oct 5;305(5933):391-7
PMID: 6353245
-
Inhibition of RNA cleavage but not polyadenylation by a point mutation in mRNA 3' consensus sequence AAUAAA.
Nature. 1983 Oct 13-19;305(5935):600-5
PMID: 6194440
-
Cloning and sequence analysis of two monkey metallothionein cDNAs.
Gene. 1983 Sep;24(1):137-46
PMID: 6414888
-
The expression in yeast of the Escherichia coli galK gene on CYC1::galK fusion plasmids.
Gene. 1983 Nov;25(2-3):249-62
PMID: 6198241
-
Primary structure and transcription of an amplified genetic locus: the CUP1 locus of yeast.
Proc Natl Acad Sci U S A. 1984 Jan;81(2):337-41
PMID: 6364141
-
Interaction of super-repressible and dominant constitutive mutations for the synthesis of galactose pathway enzymes in Saccharomyces cerevisiae.
Mol Gen Genet. 1977 Apr 29;152(3):137-44
PMID: 327270