Properties and heterologous expression of the glucose transporter GHT1 from Schizosaccharomyces pombe.
Genomic DNA of the Schizosaccharomyces pombe glucose transporter, GHT1, was obtained by complementation of the glucose transport deficient Sz. pombe strain YGS-5. Here we describe the GHT1 gene that encodes a protein of 565 amino acids with a corresponding molecular mass of 62.5 kDa. This eukaryotic glucose transporter contains 12 putative transmembrane segments and is homologous to the HXT multigene family of S. cerevisiae with several amino acid motifs of this sugar transporter family. It is also homologous to other sugar carriers from human, mouse and Escherichia coli. The function of the Ght1 protein as a glucose transporter was proved both by homologous and heterologous expression in the Sz. pombe mutant YGS-5 and in the S. cerevisiae hxt mutant RE700A, respectively. Both transformed yeast strains transported D-glucose with substrate specificity similar to that in Sz. pombe wild-type cells. Moreover, the cells of the two transformed yeast strains accumulated 2-deoxy-D-glucose, a non-metabolizable D-glucose analogue, with an efficiency similar to Sz. pombe wild-type cells. The ability of the S. cerevisiae mutant RE700A to accumulate 2DG in an delta mu H+ dependent manner after transformation with GHT1 provides evidence that the Sz. pombe transporter catalyses an energy-dependent uptake of glucose.
Lichtenberg-Fraté
H
H
Botanisches Institut der Universität Bonn.
Näschen
T
T
Heiland
S
S
Höfer
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D051792:Basic Helix-Loop-Helix Proteins; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D003847:Deoxyglucose / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D015966:Gene Expression Regulation, Fungal; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D015394:Molecular Structure; D009004:Monosaccharide Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000235:genetics; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014157:Transcription Factors / Q000235:genetics; D014170:Transformation, Genetic
D051792:Basic Helix-Loop-Helix Proteins; D003847:Deoxyglucose; D005947:Glucose; D009004:Monosaccharide Transport Proteins; D014157:Transcription Factors; C095572:Hand1 protein, mouse