tailieunhanh - Báo cáo Y học: Interaction of the GTS1 gene product with glyceraldehyde3-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. . | Eur. J. Biochem. 269 3560-3569 2002 FEBS 2002 doi 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 Weidong Liu Jinqing Wang Kazuhiro Mitsui Hua Shen and Kunio Tsurugi Department of Biochemistry Yamanashi Medical University Japan 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 glyceralde-hyde-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. Keywords continuous culture glyceraldehyde-3-phosphate dehydrogenase Gts1p metabolic oscillation yeast. Ultradian cycles with a period shorter than 24 h oscillations of the glycolytic pathway were induced after addition of glucose by inhibiting mitochondrial respiration with cyanide in cell suspensions 1-3 or cell extracts 4 of yeast with a periodicity of 1-2 min as monitored by measuring the level of NAD P H reviewed in 5 . The .