Loss of regulators of vacuolar ATPase function and ceramide synthesis results in multidrug sensitivity in schizosaccharomyces pombe.

2.50
Hdl Handle:
http://hdl.handle.net/10541/59073
Title:
Loss of regulators of vacuolar ATPase function and ceramide synthesis results in multidrug sensitivity in schizosaccharomyces pombe.
Authors:
Dawson, Keren; Toone, W Mark; Jones, Nic; Wilkinson, Caroline R M
Abstract:
We undertook a screen to isolate determinants of drug resistance in fission yeast and identified two genes that, when mutated, result in sensitivity to a range of structurally unrelated compounds, some of them commonly used in the clinic. One gene, rav1, encodes the homologue of a budding yeast protein which regulates the assembly of the vacuolar ATPase. The second gene, lac1, encodes a homologue of genes that are required for ceramide synthesis. Both mutants are sensitive to the chemotherapeutic agent doxorubicin, and using the naturally fluorescent properties of this compound, we found that both rav1 and lac1 mutations result in an increased accumulation of the drug in cells. The multidrug-sensitive phenotype of rav1 mutants can be rescued by up-regulation of the lag1 gene which encodes a homologue of lac1, whereas overexpression of either lac1 or lag1 confers multidrug resistance on wild-type cells. These data suggest that changing the amount of ceramide synthase activity in cells can influence innate drug resistance. The function of Rav1 appears to be conserved, as we show that SpRav1 is part of a RAVE-like complex in fission yeast and that loss of rav1 results in defects in vacuolar (H(+))-ATPase activity. Thus, we conclude that loss of normal V-ATPase function results in an increased sensitivity of Schizosaccharomyces pombe cells to drugs. The rav1 and lac1 genes are conserved in both higher eukaryotes and various pathogenic fungi. Thus, our data could provide the basis for strategies to sensitize tumor cells or drug-resistant pathogenic fungi to drugs.
Affiliation:
Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester M20 4BX, United Kingdom.
Citation:
Loss of regulators of vacuolar ATPase function and ceramide synthesis results in multidrug sensitivity in schizosaccharomyces pombe. 2008, 7 (6):926-37 Eukaryotic Cell
Journal:
Eukaryotic Cell
Issue Date:
Jun-2008
URI:
http://hdl.handle.net/10541/59073
DOI:
10.1128/EC.00037-08
PubMed ID:
18441123
Type:
Article
Language:
en
ISSN:
1535-9786
Appears in Collections:
All Paterson Institute for Cancer Research; Cell Regulation

Full metadata record

DC FieldValue Language
dc.contributor.authorDawson, Keren-
dc.contributor.authorToone, W Mark-
dc.contributor.authorJones, Nic-
dc.contributor.authorWilkinson, Caroline R M-
dc.date.accessioned2009-04-02T15:53:03Z-
dc.date.available2009-04-02T15:53:03Z-
dc.date.issued2008-06-
dc.identifier.citationLoss of regulators of vacuolar ATPase function and ceramide synthesis results in multidrug sensitivity in schizosaccharomyces pombe. 2008, 7 (6):926-37 Eukaryotic Cellen
dc.identifier.issn1535-9786-
dc.identifier.pmid18441123-
dc.identifier.doi10.1128/EC.00037-08-
dc.identifier.urihttp://hdl.handle.net/10541/59073-
dc.description.abstractWe undertook a screen to isolate determinants of drug resistance in fission yeast and identified two genes that, when mutated, result in sensitivity to a range of structurally unrelated compounds, some of them commonly used in the clinic. One gene, rav1, encodes the homologue of a budding yeast protein which regulates the assembly of the vacuolar ATPase. The second gene, lac1, encodes a homologue of genes that are required for ceramide synthesis. Both mutants are sensitive to the chemotherapeutic agent doxorubicin, and using the naturally fluorescent properties of this compound, we found that both rav1 and lac1 mutations result in an increased accumulation of the drug in cells. The multidrug-sensitive phenotype of rav1 mutants can be rescued by up-regulation of the lag1 gene which encodes a homologue of lac1, whereas overexpression of either lac1 or lag1 confers multidrug resistance on wild-type cells. These data suggest that changing the amount of ceramide synthase activity in cells can influence innate drug resistance. The function of Rav1 appears to be conserved, as we show that SpRav1 is part of a RAVE-like complex in fission yeast and that loss of rav1 results in defects in vacuolar (H(+))-ATPase activity. Thus, we conclude that loss of normal V-ATPase function results in an increased sensitivity of Schizosaccharomyces pombe cells to drugs. The rav1 and lac1 genes are conserved in both higher eukaryotes and various pathogenic fungi. Thus, our data could provide the basis for strategies to sensitize tumor cells or drug-resistant pathogenic fungi to drugs.en
dc.language.isoenen
dc.subjectDrug Sensitivityen
dc.subjectChemotherapyen
dc.subjectFungal Infectionen
dc.subject.meshAntifungal Agents-
dc.subject.meshCell Membrane-
dc.subject.meshDoxorubicin-
dc.subject.meshDrug Resistance, Fungal-
dc.subject.meshGene Expression Regulation, Fungal-
dc.subject.meshHot Temperature-
dc.subject.meshMutation-
dc.subject.meshSchizosaccharomyces-
dc.subject.meshSchizosaccharomyces Pombe Proteins-
dc.subject.meshSterols-
dc.subject.meshVacuolar Proton-Translocating ATPases-
dc.titleLoss of regulators of vacuolar ATPase function and ceramide synthesis results in multidrug sensitivity in schizosaccharomyces pombe.en
dc.typeArticleen
dc.contributor.departmentPaterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester M20 4BX, United Kingdom.en
dc.identifier.journalEukaryotic Cellen

Related articles on PubMed

All Items in Christie are protected by copyright, with all rights reserved, unless otherwise indicated.