The yeast Mig1 transcriptional repressor is dephosphorylated by glucose-dependent and -independent mechanisms

A yeast Saccharomyces cerevisiae Snf1 kinase, an analog of mammalian AMPK, regulates glucose derepression of genes required for utilization of alternative carbon sources through the transcriptional repressor Mig1. It has been suggested that the Glc7-Reg1 phosphatase dephosphorylates Mig1. Here we report that Mig1 is dephosphorylated by Glc7-Reg1 in an apparently glucose-dependent mechanism but also by a mechanism independent of glucose and Glc7-Reg1. In addition to serine/threonine phosphatases another process including tyrosine phosphorylation seems crucial for Mig1 regulation. Taken together, Mig1 dephosphorylation appears to be controlled in a complex manner, in line with the importance for rapid and sensitive regulation upon altered glucose concentrations in the growth medium.

[1]  Stephan Uphoff,et al.  Frequent exchange of the DNA polymerase during bacterial chromosome replication , 2017, eLife.

[2]  Timothy A. J. Haystead,et al.  Regulatory Interactions between the Reg1-Glc7 Protein Phosphatase and the Snf1 Protein Kinase , 2000, Molecular and Cellular Biology.

[3]  R. McCartney,et al.  Regulation of Snf1 Kinase , 2001, The Journal of Biological Chemistry.

[4]  P. H. Delves,et al.  MECHANISMS IN SODIUM. , 1970 .

[5]  K. Tatchell,et al.  The REG2 gene of Saccharomyces cerevisiae encodes a type 1 protein phosphatase-binding protein that functions with Reg1p and the Snf1 protein kinase to regulate growth , 1996, Molecular and cellular biology.

[6]  J. Gordon Use of vanadate as protein-phosphotyrosine phosphatase inhibitor. , 1991, Methods in enzymology.

[7]  Mark Johnston,et al.  The nuclear exportin Msn5 is required for nuclear export of the Mig1 glucose repressor of Saccharomyces cerevisiae , 1999, Current Biology.

[8]  M. Johnston,et al.  Multiple mechanisms provide rapid and stringent glucose repression of GAL gene expression in Saccharomyces cerevisiae , 1994, Molecular and cellular biology.

[9]  M. Carlson,et al.  Heterotrimer-independent regulation of activation-loop phosphorylation of Snf1 protein kinase involves two protein phosphatases , 2012, Proceedings of the National Academy of Sciences.

[10]  H. Ronne,et al.  Control of yeast GAL genes by MIG1 repressor: a transcriptional cascade in the glucose response. , 1991, The EMBO journal.

[11]  M. Leake,et al.  Superresolution imaging of single DNA molecules using stochastic photoblinking of minor groove and intercalating dyes. , 2015, Methods.

[12]  D. Crans,et al.  The chemistry and biochemistry of vanadium and the biological activities exerted by vanadium compounds. , 2004, Chemical reviews.

[13]  R. Schiestl,et al.  Frozen competent yeast cells that can be transformed with high efficiency using the LiAc/SS carrier DNA/PEG method , 2007, Nature Protocols.

[14]  M. Leake,et al.  Single-Molecule Narrow-Field Microscopy of Protein-DNA Binding Dynamics in Glucose Signal Transduction of Live Yeast Cells. , 2016, Methods in molecular biology.

[15]  S. Mathivanan,et al.  A curated compendium of phosphorylation motifs , 2007, Nature Biotechnology.

[16]  Noam Slonim,et al.  Glucose regulates transcription in yeast through a network of signaling pathways , 2009, Molecular systems biology.

[17]  N. Blom,et al.  Sequence and structure-based prediction of eukaryotic protein phosphorylation sites. , 1999, Journal of molecular biology.

[18]  M. Carlson,et al.  Glucose-regulated interaction of a regulatory subunit of protein phosphatase 1 with the Snf1 protein kinase in Saccharomyces cerevisiae. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[19]  Karin Elbing,et al.  Purification and characterization of the three Snf1-activating kinases of Saccharomyces cerevisiae. , 2006, The Biochemical journal.

[20]  M. Carlson,et al.  The GLC7 type 1 protein phosphatase is required for glucose repression in Saccharomyces cerevisiae , 1994, Molecular and cellular biology.

[21]  David Carling,et al.  Activation of yeast Snf1 and mammalian AMP-activated protein kinase by upstream kinases , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Inhibition of Bacteriophage λ protein phosphatase by organic and oxoanion inhibitors , 2002 .

[23]  D. Tzamarias,et al.  The Snf1 kinase controls glucose repression in yeast by modulating interactions between the Mig1 repressor and the Cyc8‐Tup1 co‐repressor , 2004, EMBO reports.

[24]  D. Botstein,et al.  Mutants of yeast defective in sucrose utilization. , 1981, Genetics.

[25]  I. Baranowska-Bosiacka,et al.  Biochemical and medical importance of vanadium compounds. , 2012, Acta biochimica Polonica.

[26]  Stefan Hohmann,et al.  Molecular communication: crosstalk between the Snf1 and other signaling pathways. , 2015, FEMS yeast research.

[27]  J Wu,et al.  Multiple regulatory proteins mediate repression and activation by interaction with the yeast Mig1 binding site , 1998, Yeast.

[28]  G. Wadhams,et al.  Stoichiometry and turnover in single, functioning membrane protein complexes , 2006, Nature.

[29]  M. Carlson,et al.  Repression by SSN6-TUP1 is directed by MIG1, a repressor/activator protein. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[30]  Marija Cvijovic,et al.  Yeast AMP-activated Protein Kinase Monitors Glucose Concentration Changes and Absolute Glucose Levels* , 2014, The Journal of Biological Chemistry.

[31]  Determinants of substrate recognition in nonreceptor tyrosine kinases. , 2003, Accounts of chemical research.

[32]  W. Godchaux,et al.  Fluoride inhibition of the initiation of protein synthesis in the reticulocyte lysate cell-free system. , 1975, The Journal of biological chemistry.

[33]  D. Sherratt,et al.  Stoichiometry and Architecture of Active DNA Replication Machinery in Escherichia coli , 2010, Science.

[34]  J. Eckert Diffuse hair loss and psychiatric disturbance. , 1975, Acta dermato-venereologica.

[35]  Mark C Leake,et al.  Single-Molecule Observation of DNA Replication Repair Pathways in E. coli. , 2016, Advances in experimental medicine and biology.

[36]  M. Johnston,et al.  Regulated nuclear translocation of the Mig1 glucose repressor. , 1997, Molecular biology of the cell.

[37]  R. Trumbly,et al.  In vitro characterization of the Mig1 repressor from Saccharomyces cerevisiae reveals evidence for monomeric and higher molecular weight forms , 2006, Yeast.

[38]  J. Stewart,et al.  Mutants of yeast defective in iso-1-cytochrome c. , 1974, Genetics.

[39]  M. Leake,et al.  Millisecond single-molecule localization microscopy combined with convolution analysis and automated image segmentation to determine protein concentrations in complexly structured, functional cells, one cell at a time. , 2015, Faraday discussions.

[40]  K. Shokat,et al.  Chemical Genetics: Where Genetics and Pharmacology Meet , 2007, Cell.

[41]  G. Wadhams,et al.  Millisecond timescale slimfield imaging and automated quantification of single fluorescent protein molecules for use in probing complex biological processes. , 2009, Integrative biology : quantitative biosciences from nano to macro.

[42]  Lewis Y. Geer,et al.  Analysis of phosphorylation sites on proteins from Saccharomyces cerevisiae by electron transfer dissociation (ETD) mass spectrometry , 2007, Proceedings of the National Academy of Sciences.

[43]  C. Michels,et al.  Protein phosphatase type-1 regulatory subunits Reg1p and Reg2p act as signal transducers in the glucose-induced inactivation of maltose permease in Saccharomyces cerevisiae , 2000, Molecular and General Genetics MGG.

[44]  Marcin Maziarz,et al.  Springing into Action: Reg2 Negatively Regulates Snf1 Protein Kinase and Facilitates Recovery from Prolonged Glucose Starvation in Saccharomyces cerevisiae , 2016, Applied and Environmental Microbiology.

[45]  Chao Zhang,et al.  A Chemical Genomics Study Identifies Snf1 as a Repressor of GCN4 Translation* , 2008, Journal of Biological Chemistry.

[46]  M. Carlson,et al.  Repression by SSN 6-TUP 1 is directed by MIG 1 , a repressor / activator protein ( transcription / yeast / zinc-finger protein / glucose repression ) , 2005 .

[47]  R. McCartney,et al.  Yeast Pak1 Kinase Associates with and Activates Snf1 , 2003, Molecular and Cellular Biology.

[48]  Hans Ronne,et al.  MIG1-dependent and MIG1-independent glucose regulation of MAL gene expression in Saccharomyces cerevisiae , 1995, Current Genetics.

[49]  M. Carlson,et al.  A yeast gene that is essential for release from glucose repression encodes a protein kinase. , 1986, Science.

[50]  A. Verin,et al.  Mechanisms of sodium fluoride-induced endothelial cell barrier dysfunction: role of MLC phosphorylation. , 2001, American journal of physiology. Lung cellular and molecular physiology.

[51]  M. Matsuoka,et al.  Phosphorylation of p53 at serine 15 in A549 pulmonary epithelial cells exposed to vanadate: involvement of ATM pathway. , 2007, Toxicology and applied pharmacology.

[52]  K. Entian,et al.  Carbon Source-Dependent Phosphorylation of Hexokinase PII and Its Role in the Glucose-Signaling Response in Yeast , 1998, Molecular and Cellular Biology.

[53]  A. Nairn,et al.  Protein phosphatases: recent progress. , 1991, Advances in second messenger and phosphoprotein research.

[54]  Pilar Herrero,et al.  Hxk2 Regulates the Phosphorylation State of Mig1 and Therefore Its Nucleocytoplasmic Distribution* , 2007, Journal of Biological Chemistry.

[55]  J. Scott,et al.  Mammalian AMP-activated protein kinase is homologous to yeast and plant protein kinases involved in the regulation of carbon metabolism. , 1994, The Journal of biological chemistry.

[56]  Q. Wei,et al.  Binding of vanadium (IV) to the phosphatase calcineurin , 1995, FEBS letters.

[57]  Karen M. Arndt,et al.  Access Denied: Snf1 Activation Loop Phosphorylation Is Controlled by Availability of the Phosphorylated Threonine 210 to the PP1 Phosphatase* , 2008, Journal of Biological Chemistry.