Abstract
The uptake of 6-deoxyglucose was measured in wild-type Saccharomyces cerevisiae, in a double mutant strain lacking activity for hexokinases A and B (hxkl hxk2), in a triple mutant strain lacking activity for both hexokinases and glucokinase (hxkl hxk2 glk), and in the triple mutant with high levels of activity of single kinases restored by introduction of the cloned genes. In the wild-type strain, uptake of the glucose analog showed two components, with Km values of ca. 20 mM ("high affinity") and 250 mM ("low affinity"), respectively. The double mutant also had high- and low-affinity components, but the triple mutant showed only low-affinity uptake. Reintroduction of the single kinases to the triple mutant restored high-affinity uptake. (Other experiments on 6-deoxyglucose uptake are also presented, including the apparent use of the galactose transport system when induced.) These results show that the recent implication of the kinases in transport of glucose (L.F. Bisson and D.G. Fraenkel, Proc. Natl. Acad. Sci. U.S.A. 80:1730-1734, 1983) applies equally to the nonmetabolized analog 6-deoxyglucose and suggests that the role of the kinases in transport is not merely a consequence of metabolism of the transported compound.
Full text
PDF





Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bisson L. F., Fraenkel D. G. Involvement of kinases in glucose and fructose uptake by Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1730–1734. doi: 10.1073/pnas.80.6.1730. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
- CIRILLO V. P. Mechanism of glucose transport across the yeast cell membrane. J Bacteriol. 1962 Sep;84:485–491. doi: 10.1128/jb.84.3.485-491.1962. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cirillo V. P. Relationship between sugar structure and competition for the sugar transport system in Bakers' yeast. J Bacteriol. 1968 Feb;95(2):603–611. doi: 10.1128/jb.95.2.603-611.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Franzusoff A., Cirillo V. P. Uptake and phosphorylation of 2-deoxy-D-glucose by wild-type and single-kinase strains of Saccharomyces cerevisiae. Biochim Biophys Acta. 1982 Jun 14;688(2):295–304. doi: 10.1016/0005-2736(82)90340-6. [DOI] [PubMed] [Google Scholar]
- Gancedo J. M., Clifton D., Fraenkel D. G. Yeast hexokinase mutants. J Biol Chem. 1977 Jul 10;252(13):4443–4444. [PubMed] [Google Scholar]
- Jaspers H. T., van Steveninck J. Transport-associated phosphorylation of 2-deoxy-D-glucose in Saccharomyces fragilis. Biochim Biophys Acta. 1975 Oct 17;406(3):370–385. doi: 10.1016/0005-2736(75)90017-6. [DOI] [PubMed] [Google Scholar]
- Kawasaki G., Fraenkel D. G. Cloning of yeast glycolysis genes by complementation. Biochem Biophys Res Commun. 1982 Oct 15;108(3):1107–1122. doi: 10.1016/0006-291x(82)92114-3. [DOI] [PubMed] [Google Scholar]
- Kotyk A., Michaljanicová D., Veres K., Soukupová V. Transport of 4-deoxy- and 6-deoxy-D-glucose in baker's yeast. Folia Microbiol (Praha) 1975;20(6):496–503. doi: 10.1007/BF02891709. [DOI] [PubMed] [Google Scholar]
- Kotyk A. Properties of the sugar carrier in baker's yeast. II. Specificity of transport. Folia Microbiol (Praha) 1967;12(2):121–131. doi: 10.1007/BF02896872. [DOI] [PubMed] [Google Scholar]
- Meredith S. A., Romano A. H. Uptake and phosphorylation of 2-deoxy-D-glucose by wild type and respiration-deficient bakers' yeast. Biochim Biophys Acta. 1977 May 26;497(3):745–759. doi: 10.1016/0304-4165(77)90295-1. [DOI] [PubMed] [Google Scholar]
- Romano A. H. Facilitated diffusion of 6-deoxy-D-glucose in bakers' yeast: evidence against phosphorylation-associated transport of glucose. J Bacteriol. 1982 Dec;152(3):1295–1297. doi: 10.1128/jb.152.3.1295-1297.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Serrano R., Delafuente G. Regulatory properties of the constitutive hexose transport in Saccharomyces cerevisiae. Mol Cell Biochem. 1974 Dec 20;5(3):161–171. doi: 10.1007/BF01731379. [DOI] [PubMed] [Google Scholar]
- Walsh R. B., Kawasaki G., Fraenkel D. G. Cloning of genes that complement yeast hexokinase and glucokinase mutants. J Bacteriol. 1983 May;154(2):1002–1004. doi: 10.1128/jb.154.2.1002-1004.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]