Determination of in vivo kinetics of the starvation-induced Hxt5 glucose transporter of<i>Saccharomyces cerevisiae</i>

Buziol, Stefan; Becker, Jessica; Baumeister, Anja; Jung, Susanne; Mauch, Klaus; Reuß, Matthias; Boles, Eckhard

doi:10.1111/j.1567-1364.2002.tb00096.x

Cited by 17 publications

(14 citation statements)

References 33 publications

(79 reference statements)

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“…It is apparently regulated by the growth rate of the cells rather than by the external glucose concentration (Verwaal et al , 2002). In addition, it is upregulated during growth on ethanol and on glycerol (Greatrix & van Vuuren, 2006) and upon nitrogen and carbon starvation (Buziol et al , 2002).…”

Section: Hexose Transportmentioning

confidence: 99%

Hexose and pentose transport in ascomycetous yeasts: an overview

Leandro

Fonseca

Gonçalves

2009

FEMS Yeast Research

128

106

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The biochemical characterization of sugar uptake in yeasts started five decades ago and led to the early production of abundant kinetic and mechanistic data. However, the first accurate overview of the underlying sugar transporter genes was obtained relatively late, due mainly to the genetic complexity of hexose uptake in the model yeast Saccharomyces cerevisiae. The genomic era generated in turn a massive amount of information, allowing the identification of a multitude of putative sugar transporter and sensor-encoding genes in yeast genomes, many of which are phylogenetically related. This review aims to briefly summarize our current knowledge on the biochemical and molecular features of the transporters of hexoses and pentoses in yeasts, when possible establishing links between previous kinetic studies and genomic data currently available. Emphasis is given to recent developments concerning the identification of d-xylose and l-arabinose transporter genes, which are thought to be key players in the optimization of S. cerevisiae strains for bioethanol production from lignocellulose hydrolysates.

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Section: Hexose Transportmentioning

confidence: 99%

Hexose and pentose transport in ascomycetous yeasts: an overview

Leandro

Fonseca

Gonçalves

2009

FEMS Yeast Research

128

106

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“…A family of 18 individual systems (Hxt1-17 and Gal2) and two related, signal proteins (Snf3p and Rgt2p) have shown to be involved in sugar transport [10,11]. Expression of these genes seems to be highly regulated by the glucose concentration [12][13][14][15][16], and by using knockout mutants the kinetic properties of the different transporters have been determined for glucose [17,18].…”

Section: Introductionmentioning

confidence: 98%

Modeling simultaneous glucose and xylose uptake in Saccharomyces cerevisiae from kinetics and gene expression of sugar transporters

Bertilsson

Andersson

Lidén

2007

Bioprocess Biosyst Eng

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A kinetic model for glucose and xylose co-substrate uptake in Saccharomyces cerevisiae is presented. The model couples the enzyme kinetics with the glucose-dependent genetic expression of the individual transport proteins. This novel approach implies several options for optimizing the co-substrate utilization. Interestingly, the simulations predict a maximum xylose uptake rate at a glucose concentration >0 g/L, which suggests that the genetic expressions of the considered transport proteins are of importance when optimizing the xylose uptake. This was also evident in fed-batch simulations, where a distinct optimal glucose addition rate >0 g/L x h was found. Strategies for improving the co-substrate utilization by genetic engineering of the transport systems are furthermore suggested based on simulations.

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“…To determine the glucose transport activity of rat GLUT1, the GLUT1 V69 → M allele was constitutively expressed in strain JBY15 which is unable to take up hexoses and carries an integration cassette at former HXT367 site containing the truncated, constitutive promoter of HXT7, the Kluyveromyces lactis URA3 open reading frame for counter selection and the HXT7 terminator [19,28]. The rat GLUT1 …”

Section: Construction Of Yeast Strainsmentioning

confidence: 99%

Characterisation of Mammalian GLUT Glucose Transporters in a Heterologous Yeast Expression System

Wieczorke

Dlugai²,

Krampe³

et al. 2003

Cell Physiol Biochem

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104

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We have developed a new heterologous expression system for mammalian glucose transporters. The system is based on a Saccharomyces cerevisiae strain completely deleted for all its endogenous hexose transporters and unable to take up and to grow on hexoses. To target the heterologous glucose transporters into the yeast plasma membrane in a fully active form, additional mutations had to be introduced into the hexose transport-deficient strain. Although GLUT1 was localized at the cell surface already in the parent strain, it supported uptake of glucose only in an Δhxt fgy1-1 mutant strain. Moreover, various mutations within the first half of the second predicted transmembrane helix converted GLUT1 into a form able to support uptake of glucose into yeast cells. GLUT4 was trapped in intracellular structures but became functionally expressed in the plasma membrane in Δhxt fgy1-1 fgy4X mutant strains. Glucose transport kinetics were determined with intact yeast cells by zero-trans influx measurements with a Km of 3.2 mM for human GLUT1 and of 12.6 mM for human GLUT4. Cytochalasin B inhibited these activities. Growth tests revealed that both transporter proteins are able to mediate uptake of glucose, mannose and galactose, but not of fructose. The new heterologous expression system should be a valuable tool to develop cell based high-throughput screening assays for identifying pharmaceutical compounds influencing the transporters.

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Determination of in vivo kinetics of the starvation-induced Hxt5 glucose transporter ofSaccharomyces cerevisiae

Cited by 17 publications

References 33 publications

Hexose and pentose transport in ascomycetous yeasts: an overview

Hexose and pentose transport in ascomycetous yeasts: an overview

Modeling simultaneous glucose and xylose uptake in Saccharomyces cerevisiae from kinetics and gene expression of sugar transporters

Characterisation of Mammalian GLUT Glucose Transporters in a Heterologous Yeast Expression System

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