A novel non-canonical mechanism of regulation of MST3 (mammalian Sterile20-related kinase 3)

The canonical pathway of regulation of the GCK (germinal centre kinase) III subgroup member, MST3 (mammalian Sterile20-related kinase 3), involves a caspase-mediated cleavage between N-terminal catalytic and C-terminal regulatory domains with possible concurrent autophosphorylation of the activation loop MST3(Thr178), induction of serine/threonine protein kinase activity and nuclear localization. We identified an alternative ‘non-canonical’ pathway of MST3 activation (regulated primarily through dephosphorylation) which may also be applicable to other GCKIII (and GCKVI) subgroup members. In the basal state, inactive MST3 co-immunoprecipitated with the Golgi protein GOLGA2/gm130 (golgin A2/Golgi matrix protein 130). Activation of MST3 by calyculin A (a protein serine/threonine phosphatase 1/2A inhibitor) stimulated (auto)phosphorylation of MST3(Thr178) in the catalytic domain with essentially simultaneous cis-autophosphorylation of MST3(Thr328) in the regulatory domain, an event also requiring the MST3(341–376) sequence which acts as a putative docking domain. MST3(Thr178) phosphorylation increased MST3 kinase activity, but this activity was independent of MST3(Thr328) phosphorylation. Interestingly, MST3(Thr328) lies immediately C-terminal to a STRAD (Sterile20-related adaptor) pseudokinase-like site identified recently as being involved in binding of GCKIII/GCKVI members to MO25 scaffolding proteins. MST3(Thr178/Thr328) phosphorylation was concurrent with dissociation of MST3 from GOLGA2/gm130 and association of MST3 with MO25, and MST3(Thr328) phosphorylation was necessary for formation of the activated MST3–MO25 holocomplex.

[1]  Liam J. McGuffin,et al.  The PSIPRED protein structure prediction server , 2000, Bioinform..

[2]  D. Geschwind,et al.  Inosine Augments the Effects of a Nogo Receptor Blocker and of Environmental Enrichment to Restore Skilled Forelimb Use after Stroke , 2011, The Journal of Neuroscience.

[3]  S. Hirai,et al.  YSK1, a novel mammalian protein kinase structurally related to Ste20 and SPS1, but is not involved in the known MAPK pathways , 1997, Oncogene.

[4]  Liam J. McGuffin,et al.  Intrinsic disorder prediction from the analysis of multiple protein fold recognition models , 2008, Bioinform..

[5]  T. Lu,et al.  Structures of human MST3 kinase in complex with adenine, ADP and Mn2+. , 2010, Acta crystallographica. Section D, Biological crystallography.

[6]  L. Benowitz,et al.  Mst3b, a purine-sensitive Ste20-like protein kinase, regulates axon outgrowth , 2006, Proceedings of the National Academy of Sciences.

[7]  J. Kyriakis,et al.  The GCK II and III subfamilies of the STE20 group kinases. , 2007, Frontiers in bioscience : a journal and virtual library.

[8]  I. Macara,et al.  Regulates LKB 1 Localization by Blocking Access to Importin-, and by Association with Crm 1 and Exportin-7 , 2007 .

[9]  D. Sawyer,et al.  Neuregulin-1 protects ventricular myocytes from anthracycline-induced apoptosis via erbB4-dependent activation of PI3-kinase/Akt. , 2003, Journal of molecular and cellular cardiology.

[10]  C. Volonté,et al.  Nerve growth factor-activated protein kinase N. Characterization and rapid near homogeneity purification by nucleotide affinity-exchange chromatography. , 1992, The Journal of biological chemistry.

[11]  Tohru Natsume,et al.  WNK1 Regulates Phosphorylation of Cation-Chloride-coupled Cotransporters via the STE20-related Kinases, SPAK and OSR1* , 2005, Journal of Biological Chemistry.

[12]  Chi-Ying F. Huang,et al.  Caspase Activation of Mammalian Sterile 20-like Kinase 3 (Mst3) , 2002, The Journal of Biological Chemistry.

[13]  C. Yuan,et al.  Identification and characterization of the nuclear import and export signals of the mammalian Ste20‐like protein kinase 3 , 2004, FEBS letters.

[14]  C. Yuan,et al.  Mammalian Ste20-like protein kinase 3 induces a caspase-independent apoptotic pathway. , 2010, The international journal of biochemistry & cell biology.

[15]  D. V. van Aalten,et al.  ATP and MO25α Regulate the Conformational State of the STRADα Pseudokinase and Activation of the LKB1 Tumour Suppressor , 2009, PLoS biology.

[16]  L. Greene,et al.  Differential inhibition of nerve growth factor responses by purine analogues: correlation with inhibition of a nerve growth factor- activated protein kinase , 1989, The Journal of cell biology.

[17]  Chi-Ying F. Huang,et al.  Inhibition of Cell Migration by Autophosphorylated Mammalian Sterile 20-Like Kinase 3 (MST3) Involves Paxillin and Protein-tyrosine Phosphatase-PEST* , 2006, Journal of Biological Chemistry.

[18]  J. Kyriakis,et al.  SOK1 Translocates from the Golgi to the Nucleus upon Chemical Anoxia and Induces Apoptotic Cell Death* , 2008, Journal of Biological Chemistry.

[19]  Robert Kopajtich,et al.  YSK1 is activated by the Golgi matrix protein GM130 and plays a role in cell migration through its substrate 14-3-3ζ , 2004, The Journal of cell biology.

[20]  Lynn F Ten Eyck,et al.  Crystal structure of a cAMP-dependent protein kinase mutant at 1.26A: new insights into the catalytic mechanism. , 2004, Journal of molecular biology.

[21]  E. Tham,et al.  Monophosphothreonyl extracellular signal-regulated kinases 1 and 2 (ERK1/2) are formed endogenously in intact cardiac myocytes and are enzymically active , 2011, Cellular signalling.

[22]  M. Nozaki,et al.  Molecular cloning of a novel mRNA sequence expressed in cleavage stage mouse embryos , 1993, Molecular reproduction and development.

[23]  M. Nozaki,et al.  Molecular characterization of the Drosophila Mo25 gene, which is conserved among Drosophila, mouse, and yeast. , 1996, DNA and cell biology.

[24]  E. Delpire,et al.  On the substrate recognition and negative regulation of SPAK, a kinase modulating Na+-K+-2Cl- cotransport activity. , 2010, American journal of physiology. Cell physiology.

[25]  Liam J. McGuffin,et al.  The IntFOLD server: an integrated web resource for protein fold recognition, 3D model quality assessment, intrinsic disorder prediction, domain prediction and ligand binding site prediction , 2011, Nucleic Acids Res..

[26]  R. Karas,et al.  Protein phosphatase 2 a ( PP 2 A ) binds within the oligomerization domain of striatin and regulates the phosphorylation and activation of the mammalian Ste 20-Like kinase Mst 3 , 2022 .

[27]  S. Pikkarainen,et al.  Nuclear Dbf2-related protein kinases (NDRs) in isolated cardiac myocytes and the myocardium: activation by cellular stresses and by phosphoprotein serine-/threonine-phosphatase inhibitors. , 2008, Cellular signalling.

[28]  E. Delpire.  The mammalian family of sterile 20p-like protein kinases , 2009, Pflügers Archiv - European Journal of Physiology.

[29]  C. Yuan,et al.  Mammalian Ste20-like protein kinase 3 mediates trophoblast apoptosis in spontaneous delivery , 2008, Apoptosis.

[30]  A. Moqrich,et al.  Molecular cloning and characterization of phocein, a protein found from the Golgi complex to dendritic spines. , 2001, Molecular biology of the cell.

[31]  A. Gingras,et al.  CCM3/PDCD10 Heterodimerizes with Germinal Center Kinase III (GCKIII) Proteins Using a Mechanism Analogous to CCM3 Homodimerization*♦ , 2011, The Journal of Biological Chemistry.

[32]  M. Mann,et al.  Kinase-selective enrichment enables quantitative phosphoproteomics of the kinome across the cell cycle. , 2008, Molecular cell.

[33]  D. Brautigan,et al.  Structurally different members of the okadaic acid class selectively inhibit protein serine/threonine but not tyrosine phosphatase activity. , 1992, Toxicon : official journal of the International Society on Toxinology.

[34]  Chi-Ying F. Huang,et al.  Zinc ion acts as a cofactor for serine/threonine kinase MST3 and has a distinct role in autophosphorylation of MST3. , 2005, Journal of inorganic biochemistry.

[35]  I. Macara,et al.  STRADalpha regulates LKB1 localization by blocking access to importin-alpha, and by association with Crm1 and exportin-7. , 2008, Molecular biology of the cell.

[36]  S. Pikkarainen,et al.  Regulation of Expression of the Rat Orthologue of Mouse Double Minute 2 (MDM2) by H2O2-induced Oxidative Stress in Neonatal Rat Cardiac Myocytes , 2009, The Journal of Biological Chemistry.

[37]  William Arbuthnot Sir Lane,et al.  A Mammalian Homolog of Yeast MOB1 Is Both a Member and a Putative Substrate of Striatin Family-Protein Phosphatase 2A Complexes* , 2001, The Journal of Biological Chemistry.

[38]  M. Howe,et al.  Mst3b, an Ste20-like kinase, regulates axon regeneration in mature CNS and PNS pathways , 2009, Nature Neuroscience.

[39]  B. Hemmings,et al.  Regulation of NDR Protein Kinase by Hydrophobic Motif Phosphorylation Mediated by the Mammalian Ste20-Like Kinase MST3 , 2005, Molecular and Cellular Biology.

[40]  E. Kinoshita,et al.  Phosphate-binding Tag, a New Tool to Visualize Phosphorylated Proteins*S , 2006, Molecular & Cellular Proteomics.

[41]  Martin R Larsen,et al.  TiO(2)-based phosphoproteomic analysis of the plasma membrane and the effects of phosphatase inhibitor treatment. , 2008, Journal of proteome research.

[42]  L. Greene,et al.  Cell-free detection and characterization of a novel nerve growth factor-activated protein kinase in PC12 cells. , 1987, The Journal of biological chemistry.

[43]  S. Elledge,et al.  A quantitative atlas of mitotic phosphorylation , 2008, Proceedings of the National Academy of Sciences.

[44]  Florian Gnad,et al.  Large-scale Proteomics Analysis of the Human Kinome , 2009, Molecular & Cellular Proteomics.

[45]  L. Berthiaume,et al.  Post-translational myristoylation: Fat matters in cellular life and death. , 2011, Biochimie.

[46]  T. Force,et al.  CCM3/PDCD10 stabilizes GCKIII proteins to promote Golgi assembly and cell orientation , 2010, Journal of Cell Science.

[47]  K. Ling,et al.  Identification of a Human Brain-specific Isoform of Mammalian STE20-like Kinase 3 That Is Regulated by cAMP-dependent Protein Kinase* , 2000, The Journal of Biological Chemistry.

[48]  Brian Raught,et al.  A PP2A Phosphatase High Density Interaction Network Identifies a Novel Striatin-interacting Phosphatase and Kinase Complex Linked to the Cerebral Cavernous Malformation 3 (CCM3) Protein*S , 2009, Molecular & Cellular Proteomics.

[49]  J. Blenis,et al.  Cloning and Characterization of a Human STE20-like Protein Kinase with Unusual Cofactor Requirements* , 1997, The Journal of Biological Chemistry.

[50]  Kei Sakamoto,et al.  LKB1-dependent signaling pathways. , 2006, Annual review of biochemistry.

[51]  D. Alessi,et al.  The WNK1 and WNK4 protein kinases that are mutated in Gordon's hypertension syndrome phosphorylate and activate SPAK and OSR1 protein kinases. , 2005, The Biochemical journal.

[52]  Hans Clevers,et al.  MO25α/β interact with STRADα/β enhancing their ability to bind, activate and localize LKB1 in the cytoplasm , 2003 .

[53]  D. Alessi,et al.  MO 25 is a master regulator of SPAK / OSR 1 and MST 3 / MST 4 / YSK 1 protein kinases , 2011 .

[54]  F. Denizot,et al.  A novel calmodulin-binding protein, belonging to the WD-repeat family, is localized in dendrites of a subset of CNS neurons , 1996, The Journal of cell biology.

[55]  N. Nakamura.  Emerging new roles of GM130, a cis-Golgi matrix protein, in higher order cell functions. , 2010, Journal of pharmacological sciences.

[56]  W. Sessa,et al.  CCM 3 signaling through sterile 20 – like kinases plays an essential role during zebrafish cardiovascular development and cerebral cavernous malformations , 2010 .

[57]  S. Knapp,et al.  Structural Comparison of Human Mammalian Ste20-Like Kinases , 2010, PloS one.

[58]  Marion Benoist,et al.  The striatin family: A new signaling platform in dendritic spines , 2006, Journal of Physiology-Paris.

[59]  R. Espinosa,et al.  Cloning and Characterization of MST4, a Novel Ste20-like Kinase* , 2001, The Journal of Biological Chemistry.

[60]  K. J. Monty,et al.  Determination of molecular weights and frictional ratios of proteins in impure systems by use of gel filtration and density gradient centrifugation. Application to crude preparations of sulfite and hydroxylamine reductases. , 1966, Biochimica et biophysica acta.

[61]  S. Kateriya,et al.  WD-40 repeat protein SG2NA has multiple splice variants with tissue restricted and growth responsive properties. , 2008, Gene.

[62]  R. Karas,et al.  Protein phosphatase 2a (PP2A) binds within the oligomerization domain of striatin and regulates the phosphorylation and activation of the mammalian Ste20-Like kinase Mst3 , 2011, BMC Biochemistry.

[63]  M. Mumby,et al.  Identification of Phosphoproteins and Their Phosphorylation Sites in the WEHI-231 B Lymphoma Cell Line* , 2004, Molecular & Cellular Proteomics.

[64]  Troels Z. Kristiansen,et al.  Cloning of MASK, a Novel Member of the Mammalian Germinal Center Kinase III Subfamily, with Apoptosis-inducing Properties* , 2002, The Journal of Biological Chemistry.

[65]  E. Tournier-Lasserve,et al.  Recent insights into cerebral cavernous malformations: the molecular genetics of CCM , 2010, The FEBS journal.

[66]  D. Alessi,et al.  MO25 is a master regulator of SPAK/OSR1 and MST3/MST4/YSK1 protein kinases , 2011, The EMBO journal.

[67]  Susan S. Taylor,et al.  Hydrophobic Motif Phosphorylation Is Not Required for Activation Loop Phosphorylation of p70 Ribosomal Protein S6 Kinase 1 (S6K1)* , 2011, The Journal of Biological Chemistry.

[68]  D. Alessi,et al.  The WNK 1 and WNK 4 protein kinases that are mutated in Gordon ’ s hypertension syndrome phosphorylate and activate SPAK and OSR 1 protein kinases , 2005 .

[69]  W. Sessa,et al.  CCM3 signaling through sterile 20-like kinases plays an essential role during zebrafish cardiovascular development and cerebral cavernous malformations. , 2010, The Journal of clinical investigation.

[70]  V. Sukhatme,et al.  Alpha- and beta-adrenergic stimulation induces distinct patterns of immediate early gene expression in neonatal rat myocardial cells. fos/jun expression is associated with sarcomere assembly; Egr-1 induction is primarily an alpha 1-mediated response. , 1990, The Journal of biological chemistry.

[71]  Norinobu M. Watanabe,et al.  The Ste20 group kinases as regulators of MAP kinase cascades. , 2001, Trends in cell biology.

[72]  Edward L. Huttlin,et al.  A Tissue-Specific Atlas of Mouse Protein Phosphorylation and Expression , 2010, Cell.

[73]  L. Kedes,et al.  Proteasome-Mediated Degradation of the Coactivator p300 Impairs Cardiac Transcription , 2000, Molecular and Cellular Biology.

[74]  Susan S. Taylor,et al.  Enhanced Dephosphorylation of cAMP-dependent Protein Kinase by Oxidation and Thiol Modification* , 2005, Journal of Biological Chemistry.

[75]  Liam J. McGuffin,et al.  FunFOLD: an improved automated method for the prediction of ligand binding residues using 3D models of proteins , 2011, BMC Bioinformatics.

[76]  Wei Wu,et al.  Mammalian sterile 20-like kinase 3 (MST3) mediates oxidative-stress-induced cell death by modulating JNK activation. , 2009, Bioscience reports.

[77]  Liam J. McGuffin,et al.  Rapid model quality assessment for protein structure predictions using the comparison of multiple models without structural alignments , 2010, Bioinform..

[78]  J. Bonventre,et al.  Activation of a human Ste20‐like kinase by oxidant stress defines a novel stress response pathway. , 1996, The EMBO journal.

[79]  Jennifer A. Prescher,et al.  Rapid detection, discovery, and identification of post‐translationally myristoylated proteins during apoptosis using a bio‐orthogonal azidomyristate analog , 2008, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.