Differential post‐transcriptional regulation of yeast mRNAs in response to high and low glucose concentrations

Glucose regulates yeast gene expression at both transcriptional and post‐transcriptional levels. Glucose strongly represses the transcription of the gluconeogenic genes, FBP1 and PCK1, and accelerates the degradation of their mRNAs. Together these mechanisms are responsible for the rapid decrease in gluconeogenic enzyme synthesis when yeast cells switch to glycolytic metabolism. In this study, we show that accelerated gluconeogenic mRNA degradation can be triggered by low concentrations of glucose (<0.02%). This sets the FBP1 and PCK1 mRNAs apart from other glucose‐sensitive mRNAs, such as the Ip mRNA, which only responds to high glucose concentrations (>1%). We also show that accelerated gluconeogenic mRNA degradation is co‐ordinated with transcriptional repression by common signalling components that include sugar kinases and Ras‐cAMP signalling. Furthermore, the ability of the low glucose signal to trigger accelerated gluconeogenic mRNA degradation depends upon the low glucose sensor, Snf3p, but not on the high glucose sensor, Rgt2p. Also, this response is influenced by reg1 and ume5 mutations, but not by grr1 or rgt1 mutations. Our data suggest that several signalling pathways co‐ordinate differential post‐transcriptional and transcriptional responses in yeast, depending upon the amount of glucose available in the medium.

[1]  J. Gancedo,et al.  Phosphorylation and inactivation of yeast fructose-bisphosphatase in vivo by glucose and by proton ionophores. A possible role for cAMP. , 2005, European journal of biochemistry.

[2]  B. Carter,et al.  5′‐Secondary structure formation, in constrast to a short string of non‐preferred codons, inhibits the translation of the pyruvate kinase mRNA in yeast , 1989, Yeast.

[3]  The AMP‐Activated Protein Kinase , 1997 .

[4]  Mike Tyers,et al.  F-Box Proteins Are Receptors that Recruit Phosphorylated Substrates to the SCF Ubiquitin-Ligase Complex , 1997, Cell.

[5]  A. Feinberg,et al.  A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. , 1983, Analytical biochemistry.

[6]  T. A. Brown,et al.  A rapid and simple method for preparation of RNA from Saccharomyces cerevisiae. , 1990, Nucleic acids research.

[7]  I. Scheffler,et al.  Cloning and characterization of the iron-sulfur subunit gene of succinate dehydrogenase from Saccharomyces cerevisiae. , 1990, The Journal of biological chemistry.

[8]  D. Fraenkel,et al.  Saccharomyces carlsbergensis fdp mutant and futile cycling of fructose 6-phosphate , 1982, Molecular and cellular biology.

[9]  H. Ronne.  Glucose repression in fungi. , 1995, Trends in genetics : TIG.

[10]  J. Londesborough,et al.  Purification of trehalose synthase from baker's yeast. Its temperature-dependent activation by fructose 6-phosphate and inhibition by phosphate. , 1993, European journal of biochemistry.

[11]  J. Thevelein,et al.  Activation of trehalase during growth induction by nitrogen sources in the yeast Saccharomyces cerevisiae depends on the free catalytic subunits of camp‐dependent protein kinase, but not on functional ras proteins , 1994, Yeast.

[12]  A. Brown,et al.  Yeast glycolytic mRNAs are differentially regulated , 1991, Molecular and cellular biology.

[13]  Mark Johnston,et al.  Grr1 of Saccharomyces cerevisiae is connected to the ubiquitin proteolysis machinery through Skp1: coupling glucose sensing to gene expression and the cell cycle , 1997, The EMBO journal.

[14]  S. Wölfl,et al.  Two glucose transporters in Saccharomyces cerevisiae are glucose sensors that generate a signal for induction of gene expression. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  K. Tatchell,et al.  SRA5 encodes the low-Km cyclic AMP phosphodiesterase of Saccharomyces cerevisiae , 1988, Molecular and cellular biology.

[16]  R. Young,et al.  A kinase–cyclin pair in the RNA polymerase II holoenzyme , 1995, Nature.

[17]  Mark Johnston,et al.  Glucose sensing and signaling by two glucose receptors in the yeast Saccharomyces cerevisiae , 1998, The EMBO journal.

[18]  A. Brown,et al.  The levels of yeast gluconeogenic mRNAs respond to environmental factors. , 1994, European journal of biochemistry.

[19]  J. Gancedo,et al.  Turnover of yeast fructose-bisphosphatase in different metabolic conditions. , 1980, European journal of biochemistry.

[20]  M. Carlson,et al.  Cyclin-dependent protein kinase and cyclin homologs SSN3 and SSN8 contribute to transcriptional control in yeast. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. François,et al.  The mechanism by which glucose increases fructose 2,6-bisphosphate concentration in Saccharomyces cerevisiae. A cyclic-AMP-dependent activation of phosphofructokinase 2. , 1984, European journal of biochemistry.

[22]  J. Gancedo.  Yeast Carbon Catabolite Repression , 1998, Microbiology and Molecular Biology Reviews.

[23]  Carlos Gancedo,et al.  Trehalose‐6‐phosphate, a new regulator of yeast glycolysis that inhibits hexokinases , 1993, FEBS letters.

[24]  M. Wigler,et al.  Rigorous feedback control of cAMP levels in Saccharomyces cerevisiae. , 1987, Genes & development.

[25]  H. Boedtker,et al.  RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. , 1977, Biochemistry.

[26]  M. Carlson,et al.  REG1 binds to protein phosphatase type 1 and regulates glucose repression in Saccharomyces cerevisiae. , 1995, The EMBO journal.

[27]  D. Hardie,et al.  The AMP-activated protein kinase--fuel gauge of the mammalian cell? , 1997, European journal of biochemistry.

[28]  S. Peltz,et al.  Measurement of mRNA decay rates in Saccharomyces cerevisiae. , 1991, Methods in enzymology.

[29]  M. Johnston,et al.  Two different repressors collaborate to restrict expression of the yeast glucose transporter genes HXT2 and HXT4 to low levels of glucose , 1996, Molecular and cellular biology.

[30]  M. Carlson,et al.  Functional Relationships of Srb10-Srb11 Kinase, Carboxy-Terminal Domain Kinase CTDK-I, and Transcriptional Corepressor Ssn6-Tup1 , 1998, Molecular and Cellular Biology.

[31]  J M Thevelein,et al.  Studies on the mechanism of the glucose-induced cAMP signal in glycolysis and glucose repression mutants of the yeast Saccharomyces cerevisiae. , 1988, European journal of biochemistry.

[32]  K. W. van de Poll,et al.  Isolation of a Regulatory Mutant of Fructose-1, 6-diphosphatase in Saccharomyces carlsbergensis , 1974, Journal of bacteriology.

[33]  J. Gancedo,et al.  Inactivation of yeast fructose-1,6-bisphosphatase. In vivo phosphorylation of the enzyme. , 1982, The Journal of biological chemistry.

[34]  J. Broach,et al.  RAS genes in Saccharomyces cerevisiae: signal transduction in search of a pathway. , 1991, Trends in genetics : TIG.

[35]  M. Johnston,et al.  Three different regulatory mechanisms enable yeast hexose transporter (HXT) genes to be induced by different levels of glucose , 1995, Molecular and cellular biology.

[36]  I. Purvis,et al.  The relationship between mRNA stability and length in Saccharomyces cerevisiae. , 1986, Nucleic acids research.

[37]  J. Thevelein.  Signal transduction in yeast , 1994, Yeast.

[38]  J. Gancedo,et al.  Regulation of yeast fructose‐1,6‐bisphosphatase in strains containing multicopy plasmids coding for this enzyme , 1988, FEBS letters.

[39]  M. Tyers,et al.  Inhibition of G1 cyclin activity by Ras/cAMP pathway in yeast , 1994 .

[40]  Mark Johnston,et al.  Function and Regulation of Yeast Hexose Transporters , 1999, Microbiology and Molecular Biology Reviews.

[41]  R. Trumbly.  Glucose repression in the yeast Saccharomyces cerevisiae , 1992, Molecular microbiology.

[42]  Rodney Rothstein,et al.  Elevated recombination rates in transcriptionally active DNA , 1989, Cell.

[43]  Zhikang Yin,et al.  Multiple signalling pathways trigger the exquisite sensitivity of yeast gluconeogenic mRNAs to glucose , 1996, Molecular microbiology.

[44]  P. Gonçalves,et al.  Starting up yeast glycolysis. , 1998, Trends in microbiology.

[45]  J. Thevelein,et al.  The RAS-adenylate cyclase pathway and cell cycle control in , 1992 .

[46]  M. Johnston,et al.  Feasting, fasting and fermenting. Glucose sensing in yeast and other cells. , 1999, Trends in genetics : TIG.

[47]  H. Feldmann,et al.  Nucleotide sequence of the phosphoenolpyruvate carboxykinase gene from Saccharomyces cerevisiae. , 1988, Nucleic acids research.

[48]  F. Zimmermann,et al.  Different internal metabolites trigger the induction of glycolytic gene expression in Saccharomyces cerevisiae , 1995, Journal of bacteriology.

[49]  L. Alberghina,et al.  Repression of growth-regulated Gl cyclin expression by cyclic AMP in budding yeast , 1994, Nature.

[50]  E. Martegani,et al.  Molecular cloning of a gene involved in glucose sensing in the yeast Saccharomyces cerevisiae , 1993, Molecular microbiology.

[51]  C T Verrips,et al.  Glucose Repression in Saccharomyces cerevisiae Is Related to the Glucose Concentration Rather Than the Glucose Flux* , 1998, The Journal of Biological Chemistry.

[52]  R. Parker,et al.  The cis acting sequences responsible for the differential decay of the unstable MFA2 and stable PGK1 transcripts in yeast include the context of the translational start codon. , 1999, RNA.

[53]  P. Thomas,et al.  Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. , 1980, Proceedings of the National Academy of Sciences of the United States of America.

[54]  M. Johnston,et al.  Rgt1p of Saccharomyces cerevisiae, a key regulator of glucose-induced genes, is both an activator and a repressor of transcription , 1996, Molecular and cellular biology.

[55]  H. Feldmann,et al.  Molecular cloning of CIF1, a yeast gene necessary for growth on glucose , 1992, Yeast.

[56]  O. Ozier-Kalogeropoulos,et al.  A simple and efficient method for direct gene deletion in Saccharomyces cerevisiae. , 1993, Nucleic acids research.

[57]  A. Goffeau,et al.  Cyclic AMP controls the plasma membrane H+‐ATPase activity from Saccharomyces cerevisiae , 1989, FEBS letters.

[58]  I. Scheffler,et al.  Genetic analysis of glucose regulation in saccharomyces cerevisiae: control of transcription versus mRNA turnover. , 1996, The EMBO journal.

[59]  Mark Johnston,et al.  5 Regulation of Carbon and Phosphate Utilization , 1992 .

[60]  I. Scheffler,et al.  Control of mRNA turnover as a mechanism of glucose repression in Saccharomyces cerevisiae , 1992, Molecular and cellular biology.

[61]  F. Zimmermann,et al.  Different signals control the activation of glycolysis in the yeast Saccharomyces cerevisiae , 1993, Yeast.

[62]  P. van der Zee,et al.  Characterization of the 56-kDa subunit of yeast trehalose-6-phosphate synthase and cloning of its gene reveal its identity with the product of CIF1, a regulator of carbon catabolite inactivation. , 1992, European journal of biochemistry.