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Glucose transport in the yeastKluyveromyces lactis

II. Transcriptional regulation of the glucose transporter geneRA1

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Summary

TheRAG1 gene encodes a membrane protein involved in the low-affinity glucose/fructose transport system of the yeastKluyveromyces lactis. Analysis of steady-state mRNA levels analysis and quantitation of expression by β-galactosidase fromRAG1-lacZ fusions assays revealed that theRAG1 gene was poorly expressed in cells grown under gluconeogenesis conditions, but was induced more than ten-fold when they were grown on various sugars. These sugars included glucose, fructose, mannose, sucrose, raffinose, as well as galactose. Nucleotide sequence and deletion analysis of the 5′ fianking region of theRAG1 gene showed that an essentialcis-acting element required for induced transcription of theRAG1 gene resided between − 615 and − 750 from the coding sequence. This region contained a 22 by purine stretch, and a pair of 11 by direct repeat sequences. The 11 by repeats harbor a CCAAT motif, a consensus sequence for binding of the yeast and mammalian HAP2/3/4-type protein complex. The transcription of theRAG1 gene was dramatically affected by three unlinked mutations,rag4, rag5 andrag8. We discuss the possible roles ofRAG4,RAG5 andRAG8 gene products in the expression of theRAG1 gene, as well as the importance of the inducibleRAG1 gene in the fermentative growth ofK. lactis.

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References

  • Baldwin SA, Henderson PJF (1989) Homologies between sugar transporters from eukaryotes and prokaryotes. Annu Rev Physiol 51:459–471

    Google Scholar 

  • Bisson LF, Fraenkel DG (1983a) Transport of 6-deoxyglucose inSaccharomyces cerevisiae. J Bacteriol 155:995–1000

    Google Scholar 

  • Bisson LF, Fraenkel DG (1983b) Involvement of kinases in glucose and fructose uptake bySaccharomyces cerevisiae. Proc Natl Acad Sci USA 80:1730–1734

    Google Scholar 

  • Bisson LF, Fraenkel DG (1984) Expression of kinase-dependent glucose uptake inSaccharomyces cerevisiae. J Bacteriol 159:1013–1017

    Google Scholar 

  • Bisson LF, Neigeborn L, Carlson M, Fraenkel DG (1987) TheSNF3 gene is required for high-affinity glucose transport inSaccharomyces cerevisiae. J Bacteriol 169:1656–1662

    Google Scholar 

  • Carlson M, Botstein D (1982) Two differentially regulated mRNAs with different 5′ ends encode secreted and intracellular forms of yeast invertase. Cell 28:145–154

    Google Scholar 

  • Celenza JL, Marshall-Carlson L, Carlson M (1988) The yeastSNF3 gene encodes a glucose transporter homologous to the mammalian protein. Proc Natl Acad Sci USA 85:2130–2134

    Google Scholar 

  • Chambers A, Stanway C, Tsang JSH, Henry Y, Kingsman AJ, Kingsman SM (1990) ARS binding factor 1 binds adjacent to RAPI at the UASs of the yeast glycolytic genes PGK andPYK1. Nucleic Acids Res 18:5393–5399

    Google Scholar 

  • Chen XJ (1987) Etude du plasmide pKD1 et développement de systèmes d'expression de genes chez la levureKluyveromyces lactis. Ph D Thesis Universitè de Paris-Sud

  • Chen XJ, Saliola M, Falcone C, Bianchi MM, Fukuhara H (1986) Sequence organization of the circular plasmid pKDI from the yeastKluyveromyces drosophilarum. Nucleic Acids Res 14:4471–4481

    Google Scholar 

  • Chen XJ, Wésolowski-Louvel M, Tanguy-Rougeau C, Fukuhara H (1991) Promoter activity associated with the left inverted terminal repeat of the killer plasmid kl from yeast. Biochimie 73:1195–1203

    Google Scholar 

  • Cheng Q, Michels C (1989) The maltose permease encoded by theMAL61 gene ofSaccharomyces cerevisiae exhibits both sequence and structural homology to other sugar transporters. Genetics 123:477–484

    Google Scholar 

  • Chodosh L, Baldwin A, Carthew R, Sharp P (1988a) Human CCAAT-binding protein has heterologous subunits. Cell 53:11–24

    Google Scholar 

  • Chodosh L, Olsen J, Hahn S, Baldwin A, Guarente L, Sharp P (1988b) A yeast and a human CCAAT-binding protein have heterologous subunits that are functionally interchangeable. Cell 53:25–35

    Google Scholar 

  • Cirillo VP (1962) Mechanism of glucose transport across the yeast cell membrane. J Bacteriol 84:485–491

    Google Scholar 

  • Dickson RC, Barr K (1983) Characterization of lactose transport inKluyveromyces lactis. J Bacteriol 154:1245–1251

    Google Scholar 

  • Does AL, Bisson LF (1989) Comparison of glucose uptake kinetics in different yeasts. J Bacteriol 171:1303–1308

    Google Scholar 

  • Dorsman JC, van Heeswijk WC, Grivell LA (1989) Yeast general transcription factor GFl: sequence requirement for binding to DNA and evolutionary conservation. Nucleic Acids Res 18:2769–2776

    Google Scholar 

  • Forsborg SL, Guarente L (1989) Identification and characterization of HAP4: A third component of the CCAAT-bound HAP2/3 heteromer. Genes Dev 3:1166–1178

    Google Scholar 

  • Goffrini P, Algeri AA, Donnini C, Wesolowski-Louvel M, Ferrero I (1989)RAG1 andRAG2: nuclear genes involved in the dependence/independence on mitochondrial respiratory function for the growth on sugars. Yeast 5:99–106

    Google Scholar 

  • Goffrini P, Wésolowski-Louvel M, Ferrero I, Fukuhara H (1990)RAG1 gene of the yeastKluyveromyces lactis codes for a sugar transporter. Nucleic Acids Res 18:5294

    Google Scholar 

  • Goffrini P, Wésolowski-Louvel M, Ferrero I (1991) A phosphoglucose isomerase gene is involved in the Rag phenotype of the yeastKluyveromyces lactis. Mol Gen Genet 228:401–409

    Google Scholar 

  • Guarente L, Mason T (1983) Heme regulates transcription of theCYC1 gene ofSaccharomyces cerevisiae via an upstream activation site. Cell 32:1279–1286

    Google Scholar 

  • Guarente L, Yocum RR, Gifford P (1982) AGAL10-CYC1 hybrid yeast promoter identifies theGAL4 regulatory region as an upstream site. Proc Natl Acad Sci USA 79:7410–7414

    Google Scholar 

  • Hong GF (1982) A systematic DNA sequencing strategy. J Mol Biol 158:539–549

    Google Scholar 

  • Keng T, Guarente L (1987) Constitutive expression of the yeastHEM1 gene is actually a composite of activation and repression. Proc Natl Acad Sci USA 84:9113–9117

    Google Scholar 

  • Kruckeberg AL, Bisson LF (1990) TheHXT2 gene ofSaccharomyces cerevisiae is required for high-affinity glucose transport. Mol Cell Biol 10:5903–5913

    Google Scholar 

  • Lang JM, Cirillo VP (1987) Glucose transport in a kinaselessSaccharomyces cerevisiae mutant. J Bacteriol 169:2932–2937

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Meilhoc E, Masson JP, Teissié J (1990) High efficiency transformation of intact yeast cells by electric field pulses. Biotechnology 8:223–227

    Google Scholar 

  • Miller J (1972) Experiments in Molecular Biology. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York

    Google Scholar 

  • Olesen JT, Guarente L (1990) The HAP2 subunit of yeast CCAAT transcriptional activator contains adjacent domains for subunit association and DNA recognition: model for the HAP2/3/4 complex. Genes Dev 4:1714–1729

    Google Scholar 

  • Olesen JT, Hahn S, Guarente L (1987) Yeast HAP2 and HAP3 activators both bind to theCYC1 upstream activation site UAS2 in an interdependent manner. Cell 51:953–961

    Google Scholar 

  • Postma E, van den Broek PJA (1990) Continuous-culture study of the regulation of glucose and fructose transport inKluyveromyces marxianus CBS 6556. J Bacteriol 172:2871–2876

    Google Scholar 

  • Royt PW (1981) Facilitated diffusion of 6-deoxy-D-glucose by the oxidative yeast,Kluyveromyces lactis. Arch Microbiol 130:87–89

    Google Scholar 

  • Royt PW, Macquillan AM (1976) Evidence of an inducible glucose transport system inKluyveromyces lactis. Biochim Biophys Acta 426:302–326

    Google Scholar 

  • Schneider JC (1989) Coordinate control of cytochrome genes. Ph D Thesis. M.I.T., Cambridge, MA

    Google Scholar 

  • Sheetz RM, Dickson RC (1981)LAC4 is the structural gene for β-galactosidase inKluyveromyces lactis. Genetics 98:729–745

    Google Scholar 

  • Szkutnicka K, Tschopp JF, Andrews L, Cirillo VP (1989) Sequence and structure of the yeast galactose transporter. J Bacteriol 171:4486–4493

    Google Scholar 

  • van Urk H, Postma E, Scheffers WA, van Dijken JP (1989) Glucose transport in Crabtree-positive and Crabtree-negative yeasts. J Gen Microbiol 135:2399–2406

    Google Scholar 

  • Wésolowski M, Algeri AA, Goffrini P, Fukuhara H (1982) Killer DNA plasmids of the yeastKluyveromyces lactis. I. Mutations affecting the killer phenotype. Curr Genet 150:137–140

    Google Scholar 

  • Wésolowski-Louvel M, Goffrini P, Ferrero I (1988) TheRAG2 gene of the yeastKluyveromyces lactis codes for a putative phosphoglucose isomerase. Nucleic Acids Res 16:8417

    Google Scholar 

  • Wésolowski-Louvel M, Goffrini P, Ferrero I, Fukuhara H (1992) Glucose transport in the yeastKluveromyces lactis. 1. Properties of an inducible low-affinity glucose transporter gene. Mol Gen Genet 233: 89–96

    Google Scholar 

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Author notes

  1. Xin Jie Chen

    Present address: Molecular and Population Genetics Group, Research School of Biological Sciences, The Australian National University, GPO Box 475, 2601, Canberra City, ACT, Australia

Authors and Affiliations

  1. Institut Curie, Section de Biologie, Centre Universitaire, Bât. 110, F-91405, Orsay Cédex, France

    Xin Jie Chen, Micheline Wésolowski-Louvel & Hiroshi Fukuhara

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  1. Xin Jie Chen
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  2. Micheline Wésolowski-Louvel
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  3. Hiroshi Fukuhara
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Communicated by C.P. Hollenberg

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Chen, X.J., Wésolowski-Louvel, M. & Fukuhara, H. Glucose transport in the yeastKluyveromyces lactis . Molec. Gen. Genet. 233, 97–105 (1992). https://doi.org/10.1007/BF00587566

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