Accumulation and release of the osmolyte glycerol is independent of the putative MIP channel Spac977.17p in Schizosaccharomyces pombe.
Schizosaccharomyces pombe accumulates glycerol as an osmotic regulatory solute in response to hyper-osmotic conditions. Upon a decrease in the external osmolarity, the intracellular glycerol levels should be adjusted in order to attain osmotic homeostasis. In this study, the patterns and kinetics of glycerol export from S. pombe were investigated. Upon a decrease in external osmolarity, glycerol was rapidly exported from cells to the external medium. The amount of glycerol released from the cells was proportional to the degree of change in the external osmolarity. The export process was well controlled and was not affected by reduced temperature. This points to S. pombe controlling glycerol export using specialized facilitating proteins as has been found in Saccharomyces cerevisiae where a MIP family channel protein Fps1p is involved. Analysis of the S. pombe databases revealed a putative transport protein (Spac977.17p) with homology to glycerol channel proteins of the MIP family. However, expression of the gene into the S. cerevisiae strain lacking a glycerol channel protein (fps1Delta mutant), did not complement the defect in glycerol export during hypo-osmotic stress. Deletion of spac977.17, did not affect glycerol accumulation or release in S. pombe. The patterns and kinetics of glycerol release in the mutant were similar to those of the wild type strains suggesting that the export process is independent of Spac977.17p, the only putative MIP family glycerol channel homologue in S. pombe. While the process of glycerol export in response to hypo-osmotic stress is similar to budding yeast, the underlying molecular mechanism in S. pombe appears distinct from that described in S. cerevisiae. Further studies are needed to elucidate the physiological role of the Spac977.17p channel.
Kayingo
Gerald
G
Department of Microbiology, University of Stellenbosch, Private Bag X1, Matieland 7602, South Africa. gerald.kayingo@yale.edu
Sirotkin
Vladimir
V
Hohmann
Stefan
S
Prior
Bernard A
BA
D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005990:Glycerol / Q000378:metabolism*; D007700:Kinetics; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D016133:Polymerase Chain Reaction; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D012568:Schizosaccharomyces / Q000378:metabolism*; D029702:Schizosaccharomyces pombe Proteins / Q000378:metabolism*; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014162:Transfection
D004271:DNA, Fungal; C093263:FPS1 protein, S cerevisiae; D008565:Membrane Proteins; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins; D005990:Glycerol