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Eukaryotic Cell, November 2008, p. 1895-1905, Vol. 7, No. 11
1535-9778/08/$08.00+0     doi:10.1128/EC.00018-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Oxygen-Dependent Transcriptional Regulator Hap1p Limits Glucose Uptake by Repressing the Expression of the Major Glucose Transporter Gene RAG1 in Kluyveromyces lactis{triangledown}

Wei-Guo Bao,1,4* Bernard Guiard,2 Zi-An Fang,1 Claudia Donnini,3 Michel Gervais,2 Flavia M. Lopes Passos,5 Iliana Ferrero,3 Hiroshi Fukuhara,3,4 and Monique Bolotin-Fukuhara1*

Université Paris Sud-11, CNRS UMR 8621, Institut de Génétique et Microbiologie, Bâtiment 400, 91405 Orsay, France,1 Centre de Génétique Moléculaire, Laboratoire propre du CNRS associé à l'Université Pierre et Marie Curie, 91198 Gif-sur-Yvette, France,2 Dipartimento di Genetica Antropologia Evoluzione, Università degli Studi di Parma, 43100 Parma, Italy,3 Institut Curie, Section de Recherche, Centre Universitaire, Bâtiment 110, 91405 Orsay, France,4 Laboratório de Fisiologia de Microrganismo, Instituto de Biotecnologia Aplicada a Agropecuária-BIOAGRO, Universidade Federal de Viçosa, 36570 000-Viçosa-MG, Brazil5

Received 15 January 2008/ Accepted 9 September 2008

The HAP1 (CYP1) gene product of Saccharomyces cerevisiae is known to regulate the transcription of many genes in response to oxygen availability. This response varies according to yeast species, probably reflecting the specific nature of their oxidative metabolism. It is suspected that a difference in the interaction of Hap1p with its target genes may explain some of the species-related variation in oxygen responses. As opposed to the fermentative S. cerevisiae, Kluyveromyces lactis is an aerobic yeast species which shows different oxygen responses. We examined the role of the HAP1-equivalent gene (KlHAP1) in K. lactis. KlHap1p showed a number of sequence features and some gene targets (such as KlCYC1) in common with its S. cerevisiae counterpart, and KlHAP1 was capable of complementing the hap1 mutation. However, the KlHAP1 disruptant showed temperature-sensitive growth on glucose, especially at low glucose concentrations. At normal temperature, 28°C, the mutant grew well, the colony size being even greater than that of the wild type. The most striking observation was that KlHap1p repressed the expression of the major glucose transporter gene RAG1 and reduced the glucose uptake rate. This suggested an involvement of KlHap1p in the regulation of glycolytic flux through the glucose transport system. The {Delta}Klhap1 mutant showed an increased ability to produce ethanol during aerobic growth, indicating a possible transformation of its physiological property to Crabtree positivity or partial Crabtree positivity. Dual roles of KlHap1p in activating respiration and repressing fermentation may be seen as a basis of the Crabtree-negative physiology of K. lactis.

* Corresponding author. Mailing address for Monique Bolotin-Fukuhara: Institut de Génétique et Microbiologie, CNRS UMR 8621, Bâtiment 400, Université Paris XI, 91405 Orsay, France. Phone: (33) 1 69 15 62 01. Fax: (33) 1 69 15 66 78. E-mail: monique.bolotin{at}igmors.u-psud.fr. Mailing address for Wei-Guo Bao: IGM, CNRS UMR 8621, Bâtiment 400, Université Paris XI, 91405 Orsay, France. Phone: (33) 1 69 15 74 95. Fax: (33) 1 69 15 72 96. E-mail: weiguo.bao{at}igmors.u-psud.fr

{triangledown} Published ahead of print on 19 September 2008.

Eukaryotic Cell, November 2008, p. 1895-1905, Vol. 7, No. 11
1535-9778/08/$08.00+0     doi:10.1128/EC.00018-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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