title: Identification of nuclear genes affecting 2-Deoxyglucose resistance in Schizosaccharomyces pombe.
abstract: 2-Deoxyglucose (2-DG) is a toxic glucose analog. To identify genes involved in 2-DG toxicity in Schizosaccharomyces pombe, we screened a wild-type overexpression library for genes which render cells 2-DG resistant. A gene we termed odr1, encoding an uncharacterized hydrolase, led to strong resistance and altered invertase expression when overexpressed. We speculate that Odr1 neutralizes the toxic form of 2-DG, similar to the Saccharomyces cerevisiae Dog1 and Dog2 phosphatases which dephosphorylate 2-DG-6-phosphate synthesized by hexokinase. In a complementary approach, we screened a haploid deletion library to identify 2-DG-resistant mutants. This screen identified the genes snf5, ypa1, pas1 and pho7 In liquid medium, deletions of these genes conferred 2-DG resistance preferentially under glucose-repressed conditions. The deletion mutants expressed invertase activity more constitutively than the control strain, indicating defects in the control of glucose repression. No S. cerevisiae orthologs of the pho7 gene is known, and no 2-DG resistance has been reported for any of the deletion mutants of the other genes identified here. Moreover, 2-DG leads to derepressed invertase activity in S. pombe, while in S. cerevisiae it becomes repressed. Taken together, these findings suggest that mechanisms involved in 2-DG resistance differ between budding and fission yeasts.
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysuru 570 006, Karnataka, India agnibhat@gmail.com.
authors.forename: Akshay
authors.initials: A
authors.lastname: Vishwanatha
authors.affiliation: Research Department of Genetics, Evolution and Environment, UCL Institute of Healthy Ageing, University College London, London WC1E 6BT, UK.
authors.forename: Charalampos
authors.initials: C
authors.lastname: Rallis
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysuru 570 006, Karnataka, India Institute of Cell Biology, University of Bern, Baltzerstrasse 4, CH-3012 Bern, Switzerland.
authors.forename: Shubha
authors.initials: S
authors.lastname: Bevkal Subramanyaswamy
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysuru 570 006, Karnataka, India.
authors.forename: Cletus Joseph Michael
authors.initials: CJ
authors.lastname: D'Souza
authors.affiliation: Research Department of Genetics, Evolution and Environment, UCL Institute of Healthy Ageing, University College London, London WC1E 6BT, UK.
authors.forename: Jürg
authors.initials: J
authors.lastname: Bähler
authors.affiliation: Department of Studies in Biochemistry, University of Mysore, Manasagangotri, Mysuru 570 006, Karnataka, India.
authors.forename: Martin Ernst
authors.initials: ME
authors.lastname: Schweingruber
mesh_terms: D000963:Antimetabolites / Q000378:metabolism*; D003847:Deoxyglucose / Q000378:metabolism*; D025141:Drug Resistance, Fungal; D017353:Gene Deletion; D015870:Gene Expression; D005800:Genes, Fungal; D005820:Genetic Testing; D012568:Schizosaccharomyces / Q000187:drug effects* / Q000235:genetics* / Q000254:growth & development
chemical_list: D000963:Antimetabolites; D003847:Deoxyglucose
keywords: 2-Deoxyglucose resistance; deletion library; fission yeast; glucose metabolism; invertase activity; odr1 overexpression