Transcriptional activation mediated by the yeast AP-1 protein is required for normal cadmium tolerance

J A Wemmie; A L Wu; K D Harshman; C S Parker; W S Moye-Rowley

doi:10.1016/S0021-9258(17)36680-2

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Transcriptional activation mediated by the yeast AP-1 protein is required for normal cadmium tolerance

Journal article

Open access

Peer reviewed

Transcriptional activation mediated by the yeast AP-1 protein is required for normal cadmium tolerance

J A Wemmie, A L Wu, K D Harshman, C S Parker and W S Moye-Rowley

The Journal of biological chemistry, Vol.269(20), pp.14690-14697

05/20/1994

DOI: 10.1016/S0021-9258(17)36680-2

PMID: 8182076

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Abstract

The yeast YAP1 gene encodes a transcriptional regulatory protein that utilizes a basic region-leucine zipper (bZip) DNA-binding domain to recognize its cognate DNA element. A synthetic reporter gene containing a SV40 AP-1 response element (ARE) cloned upstream of a TRP5 promoter-lacZ gene fusion shows yAP-1-dependent transactivation in vivo. Recent work has shown that changes in the gene dosage of this factor can dramatically alter the ability of a cell to tolerate a host of toxic agents including cadmium, cycloheximide, and sulfometuron methyl. We have focused on the YAP1-dependent cadmium resistance as cells that lack a functional YAP1 gene are hypersensitive to this metal. Deletion mapping experiments define two domains in the carboxyl-terminal region of the yAP-1 protein that are required for normal cadmium tolerance and ARE-TRP5-lacZ expression. Single amino acid substitutions in the bZip domain of yAP-1 indicate that this region is required for normal DNA binding and in vivo function of the protein. Replacement of a non-canonical asparagine with leucine in the yAP-1 leucine zipper leads to production of a defective protein. A substitution mutation in the basic domain converts this mutant protein into a dominant negative factor. The ability of yAP-1 to act as a positive regulator of transcription is required for its biological action.

Phenotype

Mutagenesis

Plasmids

Sequence Deletion

Cycloheximide - toxicity

Saccharomyces cerevisiae - genetics

Transcriptional Activation

Molecular Sequence Data

Saccharomyces cerevisiae - drug effects

Drug Resistance, Microbial - genetics

Recombinant Proteins - biosynthesis

DNA-Binding Proteins - metabolism

Recombinant Proteins - isolation & purification

Saccharomyces cerevisiae - metabolism

Gene Expression Regulation, Fungal - drug effects

Base Sequence

Genes, Fungal

Transcription, Genetic

Recombinant Proteins - metabolism

Cadmium - toxicity

Sulfonylurea Compounds - toxicity

Transcription Factors - biosynthesis

DNA Primers

DNA-Binding Proteins - isolation & purification

Blotting, Western

Transcription Factors - metabolism

beta-Galactosidase - biosynthesis

Saccharomyces cerevisiae Proteins

Transcription Factors - isolation & purification

DNA-Binding Proteins - biosynthesis

Fungal Proteins - metabolism

Details

Title: Subtitle: Transcriptional activation mediated by the yeast AP-1 protein is required for normal cadmium tolerance
Creators: J A Wemmie - Department of Physiology and Biophysics, University of Iowa, Iowa City 52242
A L Wu
K D Harshman
C S Parker
W S Moye-Rowley
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.269(20), pp.14690-14697
DOI: 10.1016/S0021-9258(17)36680-2
PMID: 8182076
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: GM49825 / NIGMS NIH HHS
Language: English
Date published: 05/20/1994
Academic Unit: Molecular Physiology and Biophysics; Psychiatry; Psychological and Brain Sciences; Iowa Neuroscience Institute; Neurosurgery; Internal Medicine
Record Identifier: 9984003425802771

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