Abstract
The correct biogenesis and localization of Pma1 at the plasma membrane is essential for yeast growth. A subset of PMA1 mutations behave as dominant negative because they produce aberrantly folded proteins that form protein aggregates, which in turn provoke the aggregation of the wild type protein. One approach to understand this dominant negative effect is to identify second-site mutations able to suppress the dominant lethal phenotype caused by those mutant alleles. We isolated and characterized two intragenic second-site suppressors of the PMA1-D378T dominant negative mutation. We present here the analysis of these new mutations that are located along the amino-terminal half of the protein and include a missense mutation, L151F, and an in-frame 12bp deletion that eliminates four residues from Cys409 to Ala412. The results show that the suppressor mutations disrupt the interaction between the mutant and wild type enzymes, and this enables the wild type Pma1 to reach the plasma membrane.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alleles*
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Amino Acid Sequence
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Cell Membrane / metabolism
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Molecular Sequence Data
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Mutant Proteins / chemistry*
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Mutant Proteins / genetics*
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Mutant Proteins / metabolism
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Mutation*
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Phenotype
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Protein Multimerization / genetics*
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Protein Structure, Quaternary
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Protein Structure, Tertiary
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Protein Transport
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Proton-Translocating ATPases / chemistry*
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Proton-Translocating ATPases / genetics*
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Proton-Translocating ATPases / metabolism
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Saccharomyces cerevisiae / cytology
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Saccharomyces cerevisiae / enzymology
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae Proteins / chemistry*
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Saccharomyces cerevisiae Proteins / genetics*
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Saccharomyces cerevisiae Proteins / metabolism
Substances
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Mutant Proteins
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Saccharomyces cerevisiae Proteins
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PMA1 protein, S cerevisiae
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Proton-Translocating ATPases
Grants and funding
This work was supported by the Spanish Ministerio de Ciencia e Innovación Grant BFU2008-04285. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.