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Mol Cell Biol. 1990 November; 10(11): 5903–5913.
PMCID: PMC361384

The HXT2 gene of Saccharomyces cerevisiae is required for high-affinity glucose transport.

Abstract

The HXT2 gene of the yeast Saccharomyces cerevisiae was identified on the basis of its ability to complement the defect in glucose transport of a snf3 mutant when present on the multicopy plasmid pSC2. Analysis of the DNA sequence of HXT2 revealed an open reading frame of 541 codons, capable of encoding a protein of Mr 59,840. The predicted protein displayed high sequence and structural homology to a large family of procaryotic and eucaryotic sugar transporters. These proteins have 12 highly hydrophobic regions that could form transmembrane domains; the spacing of these putative transmembrane domains is also highly conserved. Several amino acid motifs characteristic of this sugar transporter family are also present in the HXT2 protein. An hxt2 null mutant strain lacked a significant component of high-affinity glucose transport when under derepressing (low-glucose) conditions. However, the hxt2 null mutation did not incur a major growth defect on glucose-containing media. Genetic and biochemical analyses suggest that wild-type levels of high-affinity glucose transport require the products of both the HXT2 and SNF3 genes; these genes are not linked. Low-stringency Southern blot analysis revealed a number of other sequences that cross-hybridize with HXT2, suggesting that S. cerevisiae possesses a large family of sugar transporter genes.

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Selected References

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  • Asano T, Shibasaki Y, Kasuga M, Kanazawa Y, Takaku F, Akanuma Y, Oka Y. Cloning of a rabbit brain glucose transporter cDNA and alteration of glucose transporter mRNA during tissue development. Biochem Biophys Res Commun. 1988 Aug 15;154(3):1204–1211. [PubMed]
  • Baldwin SA, Henderson PJ. Homologies between sugar transporters from eukaryotes and prokaryotes. Annu Rev Physiol. 1989;51:459–471. [PubMed]
  • Bennetzen JL, Hall BD. Codon selection in yeast. J Biol Chem. 1982 Mar 25;257(6):3026–3031. [PubMed]
  • Birnbaum MJ, Haspel HC, Rosen OM. Cloning and characterization of a cDNA encoding the rat brain glucose-transporter protein. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5784–5788. [PubMed]
  • Bisson LF. High-affinity glucose transport in Saccharomyces cerevisiae is under general glucose repression control. J Bacteriol. 1988 Oct;170(10):4838–4845. [PMC free article] [PubMed]
  • Bisson LF, Fraenkel DG. Involvement of kinases in glucose and fructose uptake by Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1730–1734. [PubMed]
  • Bisson LF, Fraenkel DG. Expression of kinase-dependent glucose uptake in Saccharomyces cerevisiae. J Bacteriol. 1984 Sep;159(3):1013–1017. [PMC free article] [PubMed]
  • Bisson LF, Neigeborn L, Carlson M, Fraenkel DG. The SNF3 gene is required for high-affinity glucose transport in Saccharomyces cerevisiae. J Bacteriol. 1987 Apr;169(4):1656–1662. [PMC free article] [PubMed]
  • Burke RL, Tekamp-Olson P, Najarian R. The isolation, characterization, and sequence of the pyruvate kinase gene of Saccharomyces cerevisiae. J Biol Chem. 1983 Feb 25;258(4):2193–2201. [PubMed]
  • Busturia A, Lagunas R. Catabolite inactivation of the glucose transport system in Saccharomyces cerevisiae. J Gen Microbiol. 1986 Feb;132(2):379–385. [PubMed]
  • Cairns MT, Elliot DA, Scudder PR, Baldwin SA. Proteolytic and chemical dissection of the human erythrocyte glucose transporter. Biochem J. 1984 Jul 1;221(1):179–188. [PubMed]
  • Celenza JL, Marshall-Carlson L, Carlson M. The yeast SNF3 gene encodes a glucose transporter homologous to the mammalian protein. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2130–2134. [PubMed]
  • Chang YD, Dickson RC. Primary structure of the lactose permease gene from the yeast Kluyveromyces lactis. Presence of an unusual transcript structure. J Biol Chem. 1988 Nov 15;263(32):16696–16703. [PubMed]
  • Chen EY, Seeburg PH. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. [PubMed]
  • Cheng Q, Michels CA. The maltose permease encoded by the MAL61 gene of Saccharomyces cerevisiae exhibits both sequence and structural homology to other sugar transporters. Genetics. 1989 Nov;123(3):477–484. [PubMed]
  • Chou PY, Fasman GD. Prediction of the secondary structure of proteins from their amino acid sequence. Adv Enzymol Relat Areas Mol Biol. 1978;47:45–148. [PubMed]
  • Cohen P. Protein phosphorylation and hormone action. Proc R Soc Lond B Biol Sci. 1988 Jul 22;234(1275):115–144. [PubMed]
  • Davies A, Meeran K, Cairns MT, Baldwin SA. Peptide-specific antibodies as probes of the orientation of the glucose transporter in the human erythrocyte membrane. J Biol Chem. 1987 Jul 5;262(19):9347–9352. [PubMed]
  • Devereux J, Haeberli P, Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. [PMC free article] [PubMed]
  • Dobson MJ, Tuite MF, Roberts NA, Kingsman AJ, Kingsman SM, Perkins RE, Conroy SC, Fothergill LA. Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae. Nucleic Acids Res. 1982 Apr 24;10(8):2625–2637. [PMC free article] [PubMed]
  • Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. [PubMed]
  • Fukumoto H, Kayano T, Buse JB, Edwards Y, Pilch PF, Bell GI, Seino S. Cloning and characterization of the major insulin-responsive glucose transporter expressed in human skeletal muscle and other insulin-responsive tissues. J Biol Chem. 1989 May 15;264(14):7776–7779. [PubMed]
  • Fukumoto H, Seino S, Imura H, Seino Y, Eddy RL, Fukushima Y, Byers MG, Shows TB, Bell GI. Sequence, tissue distribution, and chromosomal localization of mRNA encoding a human glucose transporter-like protein. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5434–5438. [PubMed]
  • Garnier J, Osguthorpe DJ, Robson B. Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol. 1978 Mar 25;120(1):97–120. [PubMed]
  • Henderson PJ. The homologous glucose transport proteins of prokaryotes and eukaryotes. Res Microbiol. 1990 Mar-Apr;141(3):316–328. [PubMed]
  • Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. [PubMed]
  • Hill JE, Myers AM, Koerner TJ, Tzagoloff A. Yeast/E. coli shuttle vectors with multiple unique restriction sites. Yeast. 1986 Sep;2(3):163–167. [PubMed]
  • Hoffman CS, Winston F. A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene. 1987;57(2-3):267–272. [PubMed]
  • Janoshazi A, Solomon AK. Interaction among anion, cation and glucose transport proteins in the human red cell. J Membr Biol. 1989 Nov;112(1):25–37. [PubMed]
  • Jay D, Cantley L. Structural aspects of the red cell anion exchange protein. Annu Rev Biochem. 1986;55:511–538. [PubMed]
  • Kaestner KH, Christy RJ, McLenithan JC, Braiterman LT, Cornelius P, Pekala PH, Lane MD. Sequence, tissue distribution, and differential expression of mRNA for a putative insulin-responsive glucose transporter in mouse 3T3-L1 adipocytes. Proc Natl Acad Sci U S A. 1989 May;86(9):3150–3154. [PubMed]
  • Kayano T, Fukumoto H, Eddy RL, Fan YS, Byers MG, Shows TB, Bell GI. Evidence for a family of human glucose transporter-like proteins. Sequence and gene localization of a protein expressed in fetal skeletal muscle and other tissues. J Biol Chem. 1988 Oct 25;263(30):15245–15248. [PubMed]
  • Krebs EG, Beavo JA. Phosphorylation-dephosphorylation of enzymes. Annu Rev Biochem. 1979;48:923–959. [PubMed]
  • Kukuruzinska MA, Bergh ML, Jackson BJ. Protein glycosylation in yeast. Annu Rev Biochem. 1987;56:915–944. [PubMed]
  • Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982 May 5;157(1):105–132. [PubMed]
  • Landschulz WH, Johnson PF, McKnight SL. The leucine zipper: a hypothetical structure common to a new class of DNA binding proteins. Science. 1988 Jun 24;240(4860):1759–1764. [PubMed]
  • Maiden MC, Davis EO, Baldwin SA, Moore DC, Henderson PJ. Mammalian and bacterial sugar transport proteins are homologous. Nature. 1987 Feb 12;325(6105):641–643. [PubMed]
  • Marczynski GT, Schultz PW, Jaehning JA. Use of yeast nuclear DNA sequences to define the mitochondrial RNA polymerase promoter in vitro. Mol Cell Biol. 1989 Aug;9(8):3193–3202. [PMC free article] [PubMed]
  • Mueckler M, Caruso C, Baldwin SA, Panico M, Blench I, Morris HR, Allard WJ, Lienhard GE, Lodish HF. Sequence and structure of a human glucose transporter. Science. 1985 Sep 6;229(4717):941–945. [PubMed]
  • Mueckler M, Lodish HF. The human glucose transporter can insert posttranslationally into microsomes. Cell. 1986 Feb 28;44(4):629–637. [PubMed]
  • Neigeborn L, Carlson M. Genes affecting the regulation of SUC2 gene expression by glucose repression in Saccharomyces cerevisiae. Genetics. 1984 Dec;108(4):845–858. [PubMed]
  • Neigeborn L, Schwartzberg P, Reid R, Carlson M. Null mutations in the SNF3 gene of Saccharomyces cerevisiae cause a different phenotype than do previously isolated missense mutations. Mol Cell Biol. 1986 Nov;6(11):3569–3574. [PMC free article] [PubMed]
  • Parikh VS, Morgan MM, Scott R, Clements LS, Butow RA. The mitochondrial genotype can influence nuclear gene expression in yeast. Science. 1987 Jan 30;235(4788):576–580. [PubMed]
  • Ramos J, Cirillo VP. Role of cyclic-AMP-dependent protein kinase in catabolite inactivation of the glucose and galactose transporters in Saccharomyces cerevisiae. J Bacteriol. 1989 Jun;171(6):3545–3548. [PMC free article] [PubMed]
  • Rothstein RJ. One-step gene disruption in yeast. Methods Enzymol. 1983;101:202–211. [PubMed]
  • Sanger F, Nicklen S, Coulson AR. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. [PubMed]
  • Sauer N, Tanner W. The hexose carrier from Chlorella. cDNA cloning of a eucaryotic H+-cotransporter. FEBS Lett. 1989 Dec 18;259(1):43–46. [PubMed]
  • Schwelberger HG, Kohlwein SD, Paltauf F. Molecular cloning, primary structure and disruption of the structural gene of aldolase from Saccharomyces cerevisiae. Eur J Biochem. 1989 Mar 15;180(2):301–308. [PubMed]
  • Sikorski RS, Hieter P. A system of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics. 1989 May;122(1):19–27. [PubMed]
  • Southern EM. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. [PubMed]
  • Szkutnicka K, Tschopp JF, Andrews L, Cirillo VP. Sequence and structure of the yeast galactose transporter. J Bacteriol. 1989 Aug;171(8):4486–4493. [PMC free article] [PubMed]
  • Tanner W, Lehle L. Protein glycosylation in yeast. Biochim Biophys Acta. 1987 Apr 27;906(1):81–99. [PubMed]
  • van Urk H, Postma E, Scheffers WA, van Dijken JP. Glucose transport in crabtree-positive and crabtree-negative yeasts. J Gen Microbiol. 1989 Sep;135(9):2399–2406. [PubMed]
  • Wang LM, Weber DK, Johnson T, Sakaguchi AY. Supercoil sequencing using unpurified templates produced by rapid boiling. Biotechniques. 1988 Oct;6(9):839–843. [PubMed]
  • Leucine-zipper motif update. Nature. 1989 Jul 13;340(6229):103–104. [PubMed]
  • Zhang CC, Durand MC, Jeanjean R, Joset F. Molecular and genetical analysis of the fructose-glucose transport system in the cyanobacterium Synechocystis PCC6803. Mol Microbiol. 1989 Sep;3(9):1221–1229. [PubMed]

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