The glycerol channel Fps1p mediates the uptake of arsenite and antimonite in Saccharomyces cerevisiae

The Saccharomyces cerevisiae FPS1 gene encodes a glycerol channel protein involved in osmoregulation. We present evidence that Fps1p mediates influx of the trivalent metalloids arsenite and antimonite in yeast. Deletion of FPS1 improves tolerance to arsenite and potassium antimonyl tartrate. Under high osmolarity conditions, when the Fps1p channel is closed, wild‐type cells show the same degree of As(III) and Sb(III) tolerance as the fps1Δ mutant. Additional deletion of FPS1 in mutants defective in arsenite and antimonite detoxification partially suppresses their hypersensitivity to metalloid salts. Cells expressing a constitutively open form of the Fps1p channel are highly sensitive to both arsenite and antimonite. We also show by direct transport assays that arsenite uptake is mediated by Fps1p. Yeast cells appear to control the Fps1p‐mediated pathway of metalloid uptake, as expression of the FPS1 gene is repressed upon As(III) and Sb(III) addition. To our knowledge, this is the first report describing a eukaryotic uptake mechanism for arsenite and antimonite and its involvement in metalloid tolerance.

[1]  P. Bobrowicz,et al.  Arsenical - induced transcriptional activation of the yeast Saccharomyces cerevisiae ACR2 and ACR3 genes requires the presence of the ACR1 gene product , 1998 .

[2]  S. Hohmann,et al.  Characteristics of Fps1-dependent and -independent glycerol transport in Saccharomyces cerevisiae , 1997, Journal of bacteriology.

[3]  B. Rosen,et al.  Pathways of As(III) detoxification in Saccharomyces cerevisiae. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. Ouellette,et al.  New mechanisms of drug resistance in parasitic protozoa. , 1995, Annual review of microbiology.

[5]  Stefan Hohmann,et al.  Fps1p controls the accumulation and release of the compatible solute glycerol in yeast osmoregulation , 1999, Molecular microbiology.

[6]  B. Rosen,et al.  Purification and Characterization of Acr2p, theSaccharomyces cerevisiae Arsenate Reductase* , 2000, The Journal of Biological Chemistry.

[7]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[8]  K. Struhl,et al.  Yap, a novel family of eight bZIP proteins in Saccharomyces cerevisiae with distinct biological functions , 1997, Molecular and cellular biology.

[9]  B. Prior,et al.  Microbial MIP channels. , 2000, Trends in microbiology.

[10]  I. Herskowitz,et al.  Osmotic Balance Regulates Cell Fusion during Mating in Saccharomyces cerevisiae , 1997, The Journal of cell biology.

[11]  S. Silver Genes for all metals—a bacterial view of the Periodic Table , 1998, Journal of Industrial Microbiology and Biotechnology.

[12]  M. Malamy,et al.  Arsenate resistant mutants of Escherichia coli and phosphate transport. , 1970, Biochemical and biophysical research communications.

[13]  C. Forkner,et al.  ARSENIC AS A THERAPEUTIC AGENT IN CHRONIC MYELOGENOUS LEUKEMIA: PRELIMINARY REPORT , 1931 .

[14]  B. Rosen,et al.  Saccharomyces cerevisiae ACR2 gene encodes an arsenate reductase. , 1998, FEMS microbiology letters.

[15]  B. Rosen,et al.  The role of efflux in bacterial resistance to soft metals and metalloids. , 1999, Essays in biochemistry.

[16]  Wei Tang,et al.  Use of arsenic trioxide (As2O3) in the treatment of acute promyelocytic leukemia (APL): II. Clinical efficacy and pharmacokinetics in relapsed patients. , 1997, Blood.

[17]  B. Rosen Families of arsenic transporters. , 1999, Trends in microbiology.

[18]  J M Thevelein,et al.  GPD1, which encodes glycerol-3-phosphate dehydrogenase, is essential for growth under osmotic stress in Saccharomyces cerevisiae, and its expression is regulated by the high-osmolarity glycerol response pathway , 1994, Molecular and cellular biology.

[19]  P. Pandolfi,et al.  Complete remission after treatment of acute promyelocytic leukemia with arsenic trioxide. , 1998, The New England journal of medicine.

[20]  Rodney Rothstein,et al.  Elevated recombination rates in transcriptionally active DNA , 1989, Cell.

[21]  G. Fink,et al.  Transformation of yeast. , 1978, Proceedings of the National Academy of Sciences of the United States of America.

[22]  S. Harashima,et al.  Two new genes, PHO86 and PHO87, involved in inorganic phosphate uptake in Saccharomyces cerevisiae , 1996, Current Genetics.

[23]  S. Silver,et al.  Resistance to arsenic compounds in microorganisms. , 1994, FEMS microbiology reviews.

[24]  C. Rensing,et al.  Antimonite is accumulated by the glycerol facilitator GlpF in Escherichia coli , 1997, Journal of bacteriology.

[25]  A. Goffeau,et al.  Active efflux by multidrug transporters as one of the strategies to evade chemotherapy and novel practical implications of yeast pleiotropic drug resistance. , 1997, Pharmacology & therapeutics.

[26]  J M Thevelein,et al.  Fps1, a yeast member of the MIP family of channel proteins, is a facilitator for glycerol uptake and efflux and is inactive under osmotic stress. , 1995, The EMBO journal.

[27]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[28]  T. Lee,et al.  Cellular uptake of trivalent arsenite and pentavalent arsenate in KB cells cultured in phosphate-free medium. , 1996, Toxicology and applied pharmacology.

[29]  M. Kuo,et al.  Arsenic trioxide sensitivity is associated with low level of glutathione in cancer cells , 1999, British Journal of Cancer.

[30]  B. Rosen,et al.  Metalloid resistance mechanisms in prokaryotes. , 1998, Journal of biochemistry.

[31]  A. Dejean,et al.  Trivalent antimonials induce degradation of the PML-RAR oncoprotein and reorganization of the promyelocytic leukemia nuclear bodies in acute promyelocytic leukemia NB4 cells. , 1998, Blood.

[32]  F. Zimmermann,et al.  A yeast homologue of the bovine lens fibre MIP gene family complements the growth defect of a Saccharomyces cerevisiae mutant on fermentable sugars but not its defect in glucose‐induced RAS‐mediated cAMP signalling. , 1991, The EMBO journal.

[33]  J. Larghero,et al.  Arsenic trioxide and melarsoprol induce apoptosis in plasma cell lines and in plasma cells from myeloma patients. , 1999, Cancer research.

[34]  R. Vanholder,et al.  More than tenfold increase of arsenic in serum and packed cells of chronic hemodialysis patients. , 1993, American journal of nephrology.

[35]  B. Pearson,et al.  Construction of PCR‐ligated long flanking homology cassettes for use in the functional analysis of six unknown open reading frames from the left and right arms of Saccharomyces cerevisiae chromosome XV , 1998, Yeast.

[36]  Stefan Hohmann,et al.  The Yeast Glycerol 3-Phosphatases Gpp1p and Gpp2p Are Required for Glycerol Biosynthesis and Differentially Involved in the Cellular Responses to Osmotic, Anaerobic, and Oxidative Stress* , 2001, The Journal of Biological Chemistry.

[37]  C. Baes,et al.  The hydrolysis of cations , 1986 .

[38]  B. Rosen Bacterial resistance to heavy metals and metalloids , 1996, JBIC Journal of Biological Inorganic Chemistry.

[39]  Thomas Fiedler,et al.  A new efficient gene disruption cassette for repeated use in budding yeast , 1996, Nucleic Acids Res..

[40]  F. De Corte,et al.  Accuracy and applicability of the k0-standardization method , 1987 .

[41]  A. Goffeau,et al.  Isolation of Three Contiguous Genes, ACR1, ACR2 and ACR3, Involved in Resistance to Arsenic Compounds in the Yeast Saccharomyces cerevisiae , 1997, Yeast.

[42]  R. Wysocki,et al.  The Saccharomyces cerevisiae ACR3 Gene Encodes a Putative Membrane Protein Involved in Arsenite Transport* , 1997, The Journal of Biological Chemistry.

[43]  J M Thevelein,et al.  The two isoenzymes for yeast NAD+‐dependent glycerol 3‐phosphate dehydrogenase encoded by GPD1 and GPD2 have distinct roles in osmoadaptation and redox regulation , 1997, The EMBO journal.

[44]  M. Borgnia,et al.  Cellular and molecular biology of the aquaporin water channels. , 1999, Annual review of biochemistry.

[45]  A putative new membrane protein, Pho86p, in the inorganic phosphate uptake system of Saccharomyces cerevisiae. , 1996, Gene.

[46]  P. Philippsen,et al.  5 PCR-Based Gene Targeting in Saccharomyces cerevisiae , 1998 .

[47]  R. D. Gietz,et al.  4 Transformation of Yeast by the Lithium Acetate/Single-Stranded Carrier DNA/PEG Method , 1998 .

[48]  Larsson,et al.  Aquaporins and water homeostasis in plants. , 1999, Trends in plant science.

[49]  M. Malamy,et al.  Effect of arsenate on inorganic phosphate transport in Escherichia coli , 1980, Journal of bacteriology.

[50]  Markus J. Tamás,et al.  Stimulation of the yeast high osmolarity glycerol (HOG) pathway: evidence for a signal generated by a change in turgor rather than by water stress , 2000, FEBS letters.

[51]  P. Pandolfi,et al.  Comparative activity of melarsoprol and arsenic trioxide in chronic B-cell leukemia lines. , 1997, Blood.

[52]  R. D. Gietz,et al.  New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. , 1988, Gene.

[53]  E. Cao,et al.  Induction of apoptosis and inhibition of human gastric cancer MGC-803 cell growth by arsenic trioxide. , 1999, European journal of cancer.

[54]  W. H. Mager,et al.  The control of intracellular glycerol in Saccharomyces cerevisiae influences osmotic stress response and resistance to increased temperature , 2000, Molecular microbiology.

[55]  J. D. de Winde,et al.  Differential requirement of the yeast sugar kinases for sugar sensing in establishing the catabolite-repressed state. , 1996, European journal of biochemistry.

[56]  P. Pandolfi,et al.  Arsenic trioxide and melarsoprol induce programmed cell death in myeloid leukemia cell lines and function in a PML and PML-RARalpha independent manner. , 1998, Blood.