The transmembrane domain 10 of the yeast Pdr5p ABC antifungal efflux pump determines both substrate specificity and inhibitor susceptibility
We have previously shown that a S1360F mutation in transmembrane domain 10 (TMD10) of the Pdr5p ABC transporter modulates substrate specificity and simultaneously leads to a loss of FK506 inhibition. In this study, we have constructed and characterized the S1360F/A/T and T1364F/A/S mutations located...
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Veröffentlicht in: | Molecular microbiology 2000-03, Vol.35 (5), p.1255-1263 |
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description | We have previously shown that a S1360F mutation in transmembrane domain 10 (TMD10) of the Pdr5p ABC transporter modulates substrate specificity and simultaneously leads to a loss of FK506 inhibition. In this study, we have constructed and characterized the S1360F/A/T and T1364F/A/S mutations located in the hydrophilic face of the amphipatic Pdr5p TMD10. A T1364F mutation leads to a reduction in Pdr5p‐mediated azole and rhodamine 6G resistance. Like S1360F, the T1364F and T1364A mutants were nearly non‐responsive to FK506 inhibition. Most remarkably, however, the S1360A mutation increases FK506 inhibitor susceptibility, because Pdr5p–S1360A is hypersensitive to FK506 inhibition when compared with either wild‐type Pdr5p or the non‐responsive S1360F variant. Hence, the Pdr5p TMD10 determines both azole substrate specificity and susceptibility to reversal agents. This is the first demonstration of a eukaryotic ABC transporter where a single residue change causes either a loss or a gain in inhibitor susceptibility, depending on the nature of the mutational change. These results have important implications for the design of efficient reversal agents that could be used to overcome multidrug resistance mediated by ABC transporter overexpression. |
doi_str_mv | 10.1046/j.1365-2958.2000.01798.x |
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In this study, we have constructed and characterized the S1360F/A/T and T1364F/A/S mutations located in the hydrophilic face of the amphipatic Pdr5p TMD10. A T1364F mutation leads to a reduction in Pdr5p‐mediated azole and rhodamine 6G resistance. Like S1360F, the T1364F and T1364A mutants were nearly non‐responsive to FK506 inhibition. Most remarkably, however, the S1360A mutation increases FK506 inhibitor susceptibility, because Pdr5p–S1360A is hypersensitive to FK506 inhibition when compared with either wild‐type Pdr5p or the non‐responsive S1360F variant. Hence, the Pdr5p TMD10 determines both azole substrate specificity and susceptibility to reversal agents. This is the first demonstration of a eukaryotic ABC transporter where a single residue change causes either a loss or a gain in inhibitor susceptibility, depending on the nature of the mutational change. These results have important implications for the design of efficient reversal agents that could be used to overcome multidrug resistance mediated by ABC transporter overexpression.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1046/j.1365-2958.2000.01798.x</identifier><identifier>PMID: 10712705</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Science Ltd</publisher><subject>ABC transporter ; Amino Acid Sequence ; ATP-Binding Cassette Transporters - antagonists & inhibitors ; ATP-Binding Cassette Transporters - genetics ; ATP-Binding Cassette Transporters - metabolism ; Biological Transport ; Drug Resistance, Microbial - genetics ; Fungal Proteins - antagonists & inhibitors ; Fungal Proteins - genetics ; Fungal Proteins - metabolism ; Membrane Proteins - antagonists & inhibitors ; Membrane Proteins - genetics ; Membrane Proteins - metabolism ; Microbial Sensitivity Tests ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Pdr5 protein ; Pyrrole ; rhodamine 6G ; Saccharomyces cerevisiae ; Saccharomyces cerevisiae - drug effects ; Saccharomyces cerevisiae - metabolism ; Saccharomyces cerevisiae Proteins ; Sequence Homology, Amino Acid ; Substrate Specificity ; tacrolimus ; Tacrolimus - pharmacology</subject><ispartof>Molecular microbiology, 2000-03, Vol.35 (5), p.1255-1263</ispartof><rights>Copyright Blackwell Scientific Publications Ltd. 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In this study, we have constructed and characterized the S1360F/A/T and T1364F/A/S mutations located in the hydrophilic face of the amphipatic Pdr5p TMD10. A T1364F mutation leads to a reduction in Pdr5p‐mediated azole and rhodamine 6G resistance. Like S1360F, the T1364F and T1364A mutants were nearly non‐responsive to FK506 inhibition. Most remarkably, however, the S1360A mutation increases FK506 inhibitor susceptibility, because Pdr5p–S1360A is hypersensitive to FK506 inhibition when compared with either wild‐type Pdr5p or the non‐responsive S1360F variant. Hence, the Pdr5p TMD10 determines both azole substrate specificity and susceptibility to reversal agents. This is the first demonstration of a eukaryotic ABC transporter where a single residue change causes either a loss or a gain in inhibitor susceptibility, depending on the nature of the mutational change. These results have important implications for the design of efficient reversal agents that could be used to overcome multidrug resistance mediated by ABC transporter overexpression.</description><subject>ABC transporter</subject><subject>Amino Acid Sequence</subject><subject>ATP-Binding Cassette Transporters - antagonists & inhibitors</subject><subject>ATP-Binding Cassette Transporters - genetics</subject><subject>ATP-Binding Cassette Transporters - metabolism</subject><subject>Biological Transport</subject><subject>Drug Resistance, Microbial - genetics</subject><subject>Fungal Proteins - antagonists & inhibitors</subject><subject>Fungal Proteins - genetics</subject><subject>Fungal Proteins - metabolism</subject><subject>Membrane Proteins - antagonists & inhibitors</subject><subject>Membrane Proteins - genetics</subject><subject>Membrane Proteins - metabolism</subject><subject>Microbial Sensitivity Tests</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>Pdr5 protein</subject><subject>Pyrrole</subject><subject>rhodamine 6G</subject><subject>Saccharomyces cerevisiae</subject><subject>Saccharomyces cerevisiae - drug effects</subject><subject>Saccharomyces cerevisiae - metabolism</subject><subject>Saccharomyces cerevisiae Proteins</subject><subject>Sequence Homology, Amino Acid</subject><subject>Substrate Specificity</subject><subject>tacrolimus</subject><subject>Tacrolimus - pharmacology</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkcuO1DAQRS0EYpqBX0AWC3YJZaed2AsWQ4vHSDOCxSCxs5y4TLuVF7aj6f4HPhqHHiHEilXZuqduVekSQhmUDLb1m0PJqloUXAlZcgAogTVKlsdHZPNHeEw2oAQUleTfLsizGA8ArIK6ekouGDSMNyA25OfdHmkKZowDDm2uSO00GD_mWXRyNGX5hCYm-sUGMdOrdztqxuTdMn43PUXn-uVI52WYqcWEYfAjRtpOaU_j0sbsnJDGGTvvfOfTKTdb6se9b32aQmZih3PKvz6Lz8kTZ_qILx7qJfn64f3d7lNx8_nj9e7qpuhEpWTBRCPy4AZl3TmBWwmNAg5oedMqWxvgBmRjW-WYtQyBS2m3QnEmOovbCqpL8vrsO4fpx4Ix6cHnPfo-nz8tUbNGgAJWZ_DVP-BhWsKYd9NM1YKL7J0heYa6MMUY0Ok5-MGEk2ag17j0Qa-p6DUVvcalf8elj7n15YP_0g5o_2o855OBt2fg3vd4-m9jfXt7vb6qX5oapVg</recordid><startdate>200003</startdate><enddate>200003</enddate><creator>Egner, Ralf</creator><creator>Bauer, Bettina E.</creator><creator>Kuchler, Karl</creator><general>Blackwell Science Ltd</general><general>Blackwell Publishing Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>200003</creationdate><title>The transmembrane domain 10 of the yeast Pdr5p ABC antifungal efflux pump determines both substrate specificity and inhibitor susceptibility</title><author>Egner, Ralf ; Bauer, Bettina E. ; Kuchler, Karl</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5398-1575eff7e86cf5e48079020ed27b9d6a02a087db9f1dd1e0288d459215cde4303</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>ABC transporter</topic><topic>Amino Acid Sequence</topic><topic>ATP-Binding Cassette Transporters - antagonists & inhibitors</topic><topic>ATP-Binding Cassette Transporters - genetics</topic><topic>ATP-Binding Cassette Transporters - metabolism</topic><topic>Biological Transport</topic><topic>Drug Resistance, Microbial - genetics</topic><topic>Fungal Proteins - antagonists & inhibitors</topic><topic>Fungal Proteins - genetics</topic><topic>Fungal Proteins - metabolism</topic><topic>Membrane Proteins - antagonists & inhibitors</topic><topic>Membrane Proteins - genetics</topic><topic>Membrane Proteins - metabolism</topic><topic>Microbial Sensitivity Tests</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>Pdr5 protein</topic><topic>Pyrrole</topic><topic>rhodamine 6G</topic><topic>Saccharomyces cerevisiae</topic><topic>Saccharomyces cerevisiae - drug effects</topic><topic>Saccharomyces cerevisiae - metabolism</topic><topic>Saccharomyces cerevisiae Proteins</topic><topic>Sequence Homology, Amino Acid</topic><topic>Substrate Specificity</topic><topic>tacrolimus</topic><topic>Tacrolimus - pharmacology</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Egner, Ralf</creatorcontrib><creatorcontrib>Bauer, Bettina E.</creatorcontrib><creatorcontrib>Kuchler, Karl</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Egner, Ralf</au><au>Bauer, Bettina E.</au><au>Kuchler, Karl</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>The transmembrane domain 10 of the yeast Pdr5p ABC antifungal efflux pump determines both substrate specificity and inhibitor susceptibility</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>2000-03</date><risdate>2000</risdate><volume>35</volume><issue>5</issue><spage>1255</spage><epage>1263</epage><pages>1255-1263</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><abstract>We have previously shown that a S1360F mutation in transmembrane domain 10 (TMD10) of the Pdr5p ABC transporter modulates substrate specificity and simultaneously leads to a loss of FK506 inhibition. In this study, we have constructed and characterized the S1360F/A/T and T1364F/A/S mutations located in the hydrophilic face of the amphipatic Pdr5p TMD10. A T1364F mutation leads to a reduction in Pdr5p‐mediated azole and rhodamine 6G resistance. Like S1360F, the T1364F and T1364A mutants were nearly non‐responsive to FK506 inhibition. Most remarkably, however, the S1360A mutation increases FK506 inhibitor susceptibility, because Pdr5p–S1360A is hypersensitive to FK506 inhibition when compared with either wild‐type Pdr5p or the non‐responsive S1360F variant. Hence, the Pdr5p TMD10 determines both azole substrate specificity and susceptibility to reversal agents. This is the first demonstration of a eukaryotic ABC transporter where a single residue change causes either a loss or a gain in inhibitor susceptibility, depending on the nature of the mutational change. These results have important implications for the design of efficient reversal agents that could be used to overcome multidrug resistance mediated by ABC transporter overexpression.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>10712705</pmid><doi>10.1046/j.1365-2958.2000.01798.x</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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subjects | ABC transporter Amino Acid Sequence ATP-Binding Cassette Transporters - antagonists & inhibitors ATP-Binding Cassette Transporters - genetics ATP-Binding Cassette Transporters - metabolism Biological Transport Drug Resistance, Microbial - genetics Fungal Proteins - antagonists & inhibitors Fungal Proteins - genetics Fungal Proteins - metabolism Membrane Proteins - antagonists & inhibitors Membrane Proteins - genetics Membrane Proteins - metabolism Microbial Sensitivity Tests Molecular Sequence Data Mutagenesis, Site-Directed Pdr5 protein Pyrrole rhodamine 6G Saccharomyces cerevisiae Saccharomyces cerevisiae - drug effects Saccharomyces cerevisiae - metabolism Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid Substrate Specificity tacrolimus Tacrolimus - pharmacology |
title | The transmembrane domain 10 of the yeast Pdr5p ABC antifungal efflux pump determines both substrate specificity and inhibitor susceptibility |
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