Interaction of the GTS1 gene product with glyceraldehyde- 3-phosphate dehydrogenase 1 required for the maintenance of the metabolic oscillations of the yeast Saccharomyces cerevisiae.
We previously reported that GTS1 is involved in regulating ultradian oscillations of the glycolytic pathway induced by cyanide in cell suspensions as well as oscillations of energy metabolism in aerobic continuous cultures. Here, we screened a yeast cDNA library for proteins that bind to Gts1p using the yeast two-hybrid system and cloned multiple TDH cDNAs encoding the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found that the zinc-finger and dimerization sites of Gts1p were required for full ability to bind GAPDH, and Gts1ps mutated at these sites lost the ability to regulate both aerobic and unaerobic ultradian oscillations of energy metabolism. Of the three TDH genes, only TDH1 fluctuated at the mRNA level in continuous culture and its deletion resulted in the disappearance of the oscillation without any affect on growth rate. This loss of biological rhythms in the TDH1-deleted mutant was rescued by the expression of TDH1 but not of TDH2 or TDH3 under the control of the TDH1 promoter. Thus, we hypothesized that Gts1p plays a role in the regulation of metabolic oscillation by interacting with the TDH1 product, GAPDH1, in yeast.
Liu
Weidong
W
Department of Biochemistry, Yamanashi Medical University, Nakakoma, Japan.
Wang
Jinqing
J
Mitsui
Kazuhiro
K
Shen
Hua
H
Tsurugi
Kunio
K
D000963:Antimetabolites / Q000494:pharmacology; D001665:Binding Sites; D001683:Biological Clocks / Q000187:drug effects / Q000502:physiology*; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000378:metabolism; D019281:Dimerization; D004734:Energy Metabolism / Q000187:drug effects; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D018390:Gene Targeting; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D006019:Glycolysis / Q000187:drug effects; D025941:Protein Interaction Mapping; D012331:RNA, Fungal / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D012966:Sodium Cyanide / Q000494:pharmacology; D014157:Transcription Factors; D020798:Two-Hybrid System Techniques; D016335:Zinc Fingers / Q000502:physiology
D000963:Antimetabolites; D018076:DNA, Complementary; D004271:DNA, Fungal; D005656:Fungal Proteins; C097841:GTS1 protein, S cerevisiae; D012331:RNA, Fungal; D012333:RNA, Messenger; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); C465224:TDH1 protein, S cerevisiae; C465225:TDH2 protein, S cerevisiae; C465226:TDH3 protein, S cerevisiae; D012966:Sodium Cyanide