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J. Bacteriol., 06 1995, 3251-3258, Vol 177, No. 11
Copyright © 1995, American Society for Microbiology

Computer-assisted nonlinear regression analysis of the multicomponent glucose uptake kinetics of Saccharomyces cerevisiae

DM Coons, RB Boulton and LF Bisson
Department of Viticulture and Enology, University of California, Davis 95616-8749, USA.

The kinetics of glucose uptake in Saccharomyces cerevisiae are complex. An Eadie-Hofstee (rate of uptake versus rate of uptake over substrate concentration) plot of glucose uptake shows a nonlinear form typical of a multicomponent system. The nature of the constituent components is a subject of debate. It has recently been suggested that this nonlinearity is due to either a single saturable component together with free diffusion of glucose or a single constitutive component with a variable Km, rather than the action of multiple hexose transporters. Genetic data support the existence of a family of differentially regulated glucose transporters, encoded by the HXT genes. In this work, kinetic expressions and nonlinear regression analysis, based on an improved zero trans-influx assay, were used to address the nature of the components of the transport system. The results indicate that neither one component with free diffusion nor a single permease with a variable Km can explain the observed uptake rates. Results of uptake experiments, including the use of putative alternative substrates as inhibitory compounds, support the model derived from genetic analyses of a multicomponent system with at least two components, one a high- affinity carrier and the other a low-affinity carrier. This approach was extended to characterize the activity of the SNF3 protein and identify its role in the depression of high-affinity uptake. The kinetic data support a role of SNF3 as a regulatory protein that may not itself be a transporter.

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