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Journal of Bacteriology, September 2001, p. 5223-5229, Vol. 183, No. 18
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.18.5223-5229.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Feedback Regulation of Glucose Transporter Gene Transcription in Kluyveromyces lactis by Glucose Uptake

C. Milkowski,1,dagger S. Krampe,2 J. Weirich,2,Dagger V. Hasse,2,§ E. Boles,2 and K. D. Breunig1,2,*

Institut für Genetik, Martin-Luther-Universität Halle-Wittenberg, D-06099 Halle,1 and Institut für Mikrobiologie, Heinrich-Heine-Universität Düsseldorf, D-40225 Düsseldorf,2 Germany

Received 5 April 2001/Accepted 29 June 2001

In the respirofermentative yeast Kluyveromyces lactis, only a single genetic locus encodes glucose transporters that can support fermentative growth. This locus is polymorphic in wild-type isolates carrying either KHT1 and KHT2, two tandemly arranged HXT-like genes, or RAG1, a low-affinity transporter gene that arose by recombination between KHT1 and KHT2. Here we show that KHT1 is a glucose-induced gene encoding a low-affinity transporter very similar to Rag1p. Kht2p has a lower Km (3.7 mM) and a more complex regulation. Transcription is high in the absence of glucose, further induced by low glucose concentrations, and repressed at higher glucose concentrations. The response of KHT1 and KHT2 gene regulation to high but not to low concentrations of glucose depends on glucose transport. The function of either Kht1p or Kht2p is sufficient to mediate the characteristic response to high glucose, which is impaired in a kht1 kht2 deletion mutant. Thus, the KHT genes are subject to mutual feedback regulation. Moreover, glucose repression of the endogenous beta -galactosidase (LAC4) promoter and glucose induction of pyruvate decarboxylase were abolished in the kht1 kht2 mutant. These phenotypes could be partially restored by HXT gene family members from Saccharomyces cerevisiae. The results indicate that the specific responses to high but not to low glucose concentrations require a high rate of glucose uptake.

* Corresponding author. Mailing address: Institut für Genetik, Martin-Luther-Universität Halle-Wittenberg, D-06099 Halle, Germany. Phone: 49-345-5526301. Fax: 49-345-5527151. E-mail: breunig{at}genetik.uni-halle.de.

dagger Present address: Leibniz-Institut für Pflanzenbiochemie, Abteilung Sekundärstoffwechsel, 06120 Halle (Saale), Germany.

Dagger Present address: Aventis Pasteur MSD Gmbh, 69181 Leimen, Germany.

§ Present address: Institut für Vegetative Physiologie, D-50931 Köln, Germany.

Journal of Bacteriology, September 2001, p. 5223-5229, Vol. 183, No. 18
0021-9193/01/$04.00+0   DOI: 10.1128/JB.183.18.5223-5229.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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