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PMID:2068093

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Citation

Dameron, CT, Winge, DR, George, GN, Sansone, M, Hu, S and Hamer, D (1991) A copper-thiolate polynuclear cluster in the ACE1 transcription factor. Proc. Natl. Acad. Sci. U.S.A. 88:6127-31

Abstract

ACE1 is the transcriptional activator of the metallothionein (CUP1 locus) gene in Saccharomyces cerevisiae. Previous data had implicated the N-terminal domain of ACE1 as responsible for the Cu-dependent specific DNA binding. An expression system in Escherichia coli was constructed to enable the isolation of an ACE1 domain containing the DNA and Cu-binding regions. Here we report the purification and characterization of the Cu-ACE1 truncated molecule. Spectroscopic techniques showed that ACE1 contains an unusual type of DNA binding structure that is based on a polynuclear Cu(I)-cysteinyl thiolate cluster. The cluster consists of six or seven Cu(I) ions coordinated to cysteinyl thiolates in a trigonal geometry distorted from planarity. The Cu(I)-cysteine cluster of Cu-ACE1 exhibits structural properties analogous to the Cu(I)-thiolate polynuclear cluster in yeast Cu-metallothionein itself, suggesting an unusual mechanism for the evolution of this regulatory factor. The Cu cluster organizes and stabilizes the conformation of the N-terminal domain of ACE1 for specific DNA binding.

Links

PubMed PMC52035

Keywords

Amino Acid Sequence; Cloning, Molecular; Copper/analysis; Cysteine/analysis; DNA-Binding Proteins/chemistry; DNA-Binding Proteins/isolation & purification; DNA-Binding Proteins/metabolism; Electron Probe Microanalysis; Escherichia coli/genetics; Molecular Sequence Data; Protein Conformation; Recombinant Proteins/chemistry; Recombinant Proteins/isolation & purification; Recombinant Proteins/metabolism; Saccharomyces cerevisiae/genetics; Saccharomyces cerevisiae Proteins; Spectrophotometry, Ultraviolet; Transcription Factors/chemistry; Transcription Factors/isolation & purification; Transcription Factors/metabolism

Significance

Annotations

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

YEAST:CUP2

enables

GO:0005507: copper ion binding

ECO:0000314: direct assay evidence used in manual assertion

F

Seeded From UniProt

complete


See also

References

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