Control of the Glycolytic Flux in Saccharomyces cerevisiae Grown at Low Temperature: A MULTI-LEVEL ANALYSIS IN ANAEROBIC CHEMOSTAT CULTURES

@article{citeulike:1821082, abstract = {Growth temperature has a profound impact on the kinetic properties of enzymes in microbial metabolic networks. Activities of glycolytic enzymes in Saccharomyces cerevisiae were up to 7.5-fold lower when assayed at 12 {degrees}C than at 30 {degrees}C. Nevertheless, the in vivo glycolytic flux in chemostat cultures (dilution rate: 0.03 h-1) grown at these two temperatures was essentially the same. To investigate how yeast maintained a constant glycolytic flux despite the kinetic challenge imposed by a lower growth temperature, a systems approach was applied that involved metabolic flux analysis, transcript analysis, enzyme activity assays, and metabolite analysis. Expression of hexose-transporter genes was affected by the growth temperature, as indicated by differential transcription of five HXT genes and changed zero trans-influx kinetics of [14C]glucose transport. No such significant changes in gene expression were observed for any of the glycolytic enzymes. Fermentative capacity (assayed off-line at 30 {degrees}C), which was 2-fold higher in cells grown at 12 {degrees}C, was therefore probably controlled predominantly by glucose transport. Massive differences in the intracellular concentrations of nucleotides (resulting in an increased adenylate energy charge at low temperature) and glycolytic intermediates indicated a dominant role of metabolic control as opposed to gene expression in the adaptation of glycolytic enzyme activity to different temperatures. In evolutionary terms, this predominant reliance on metabolic control of a central pathway, which represents a significant fraction of the cellular protein of the organism, may be advantageous to limit the need for protein synthesis and degradation during adaptation to diurnal temperature cycles. 10.1074/jbc.M610845200}, author = {Tai, Siew L. and Daran-Lapujade, Pascale and Luttik, Marijke A. and Walsh, Michael C. and Diderich, Jasper A. and Krijger, Gerard C. and van Gulik, Walter M. and Pronk, Jack T. and Daran, Jean-Marc }, citeulike-article-id = {1821082}, doi = {10.1074/jbc.M610845200}, journal = {J. Biol. Chem.}, keywords = {axp, temperature}, month = {April}, number = {14}, pages = {10243--10251}, posted-at = {2007-10-25 14:55:23}, priority = {2}, title = {Control of the Glycolytic Flux in Saccharomyces cerevisiae Grown at Low Temperature: A MULTI-LEVEL ANALYSIS IN ANAEROBIC CHEMOSTAT CULTURES}, url = {http://dx.doi.org/10.1074/jbc.M610845200}, volume = {282}, year = {2007} }

TY - JOUR ID - citeulike:1821082 L3 - citeulike-article-id:1821082 TI - Control of the Glycolytic Flux in Saccharomyces cerevisiae Grown at Low Temperature: A MULTI-LEVEL ANALYSIS IN ANAEROBIC CHEMOSTAT CULTURES JF - J. Biol. Chem. VL - 282 IS - 14 SP - 10243 EP - 10251 N2 - Growth temperature has a profound impact on the kinetic properties of enzymes in microbial metabolic networks. Activities of glycolytic enzymes in Saccharomyces cerevisiae were up to 7.5-fold lower when assayed at 12 {degrees}C than at 30 {degrees}C. Nevertheless, the in vivo glycolytic flux in chemostat cultures (dilution rate: 0.03 h-1) grown at these two temperatures was essentially the same. To investigate how yeast maintained a constant glycolytic flux despite the kinetic challenge imposed by a lower growth temperature, a systems approach was applied that involved metabolic flux analysis, transcript analysis, enzyme activity assays, and metabolite analysis. Expression of hexose-transporter genes was affected by the growth temperature, as indicated by differential transcription of five HXT genes and changed zero trans-influx kinetics of [14C]glucose transport. No such significant changes in gene expression were observed for any of the glycolytic enzymes. Fermentative capacity (assayed off-line at 30 {degrees}C), which was 2-fold higher in cells grown at 12 {degrees}C, was therefore probably controlled predominantly by glucose transport. Massive differences in the intracellular concentrations of nucleotides (resulting in an increased adenylate energy charge at low temperature) and glycolytic intermediates indicated a dominant role of metabolic control as opposed to gene expression in the adaptation of glycolytic enzyme activity to different temperatures. In evolutionary terms, this predominant reliance on metabolic control of a central pathway, which represents a significant fraction of the cellular protein of the organism, may be advantageous to limit the need for protein synthesis and degradation during adaptation to diurnal temperature cycles. 10.1074/jbc.M610845200 KW - axp KW - temperature AU - Tai, Siew AU - Daran-Lapujade, Pascale AU - Luttik, Marijke AU - Walsh, Michael AU - Diderich, Jasper AU - Krijger, Gerard AU - van Gulik, Walter AU - Pronk, Jack AU - Daran, Jean-Marc PY - 2007/04/06/ UR - http://dx.doi.org/10.1074/jbc.M610845200 ER -