PRAK, a novel protein kinase regulated by the p38 MAP kinase

We have identified and cloned a novel serine/ threonine kinase, p38‐regulated/activated protein kinase (PRAK). PRAK is a 471 amino acid protein with 20–30% sequence identity to the known MAP kinase‐regulated protein kinases RSK1/2/3, MNK1/2 and MAPKAP‐K2/3. PRAK was found to be expressed in all human tissues and cell lines examined. In HeLa cells, PRAK was activated in response to cellular stress and proinflammatory cytokines. PRAK activity was regulated by p38α and p38β both in vitro and in vivo and Thr182 was shown to be the regulatory phosphorylation site. Activated PRAK in turn phosphorylated small heat shock protein 27 (HSP27) at the physiologically relevant sites. An in‐gel kinase assay demonstrated that PRAK is a major stress‐activated kinase that can phosphorylate small heat shock protein, suggesting a potential role for PRAK in mediating stress‐induced HSP27 phosphorylation in vivo.

[1]  R. Davis,et al.  Mitogen-activated protein kinase kinase 7 is an activator of the c-Jun NH2-terminal kinase. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[2]  J. Landry,et al.  Regulation of actin filament dynamics by p38 map kinase-mediated phosphorylation of heat shock protein 27. , 1997, Journal of cell science.

[3]  Jonathan A. Cooper,et al.  Mitogen and stress response pathways: MAP kinase cascades and phosphatase regulation in mammals and yeast. , 1995, Current opinion in cell biology.

[4]  C. Gabel,et al.  Inhibition of interleukin-1 beta production by SKF86002: evidence of two sites of in vitro activity and of a time and system dependence. , 1995, Molecular pharmacology.

[5]  C. Marshall,et al.  Activation of MAP kinase kinase is necessary and sufficient for PC12 differentiation and for transformation of NIH 3T3 cells , 1994, Cell.

[6]  Philip R. Cohen,et al.  FGF and stress regulate CREB and ATF‐1 via a pathway involving p38 MAP kinase and MAPKAP kinase‐2. , 1996, The EMBO journal.

[7]  E. Krebs,et al.  The MAPK signaling cascade , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[8]  R. Davis,et al.  The mitogen-activated protein kinase signal transduction pathway. , 1993, The Journal of biological chemistry.

[9]  David Stokoe,et al.  Identification of MAPKAP kinase 2 as a major enzyme responsible for the phosphorylation of the small mammalian heat shock proteins , 1992, FEBS letters.

[10]  J. Jongstra,et al.  LSP1 Is the Major Substrate for Mitogen-activated Protein Kinase-activated Protein Kinase 2 in Human Neutrophils* , 1997, The Journal of Biological Chemistry.

[11]  Jiahuai Han,et al.  Endotoxin induces rapid protein tyrosine phosphorylation in 70Z/3 cells expressing CD14. , 1993, The Journal of biological chemistry.

[12]  Jonathan A. Cooper,et al.  Mitogen‐activated protein kinases activate the serine/threonine kinases Mnk1 and Mnk2 , 1997, The EMBO journal.

[13]  E. Nishida,et al.  Activation and Involvement of p38 Mitogen-activated Protein Kinase in Glutamate-induced Apoptosis in Rat Cerebellar Granule Cells* , 1997, The Journal of Biological Chemistry.

[14]  J. Dixon,et al.  Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. , 1991, Analytical biochemistry.

[15]  Philip R. Cohen,et al.  Identification of anisomycin-activated kinases p45 and p55 in murine cells as MAPKAP kinase-2. , 1996, Oncogene.

[16]  E. Nishida,et al.  Activation of p38 MAP kinase pathway by erythropoietin and interleukin-3. , 1997, Blood.

[17]  Yong Jiang,et al.  Characterization of the Structure and Function of the Fourth Member of p38 Group Mitogen-activated Protein Kinases, p38δ* , 1997, The Journal of Biological Chemistry.

[18]  Thomas J. White,et al.  PCR protocols: a guide to methods and applications. , 1990 .

[19]  Philip R. Cohen,et al.  Identification of novel phosphorylation sites required for activation of MAPKAP kinase‐2. , 1995, The EMBO journal.

[20]  P. Rumsby PCR protocols—A guide to methods and applications: Edited by M. A. Innis, D. H. Gelfand, J. J. Sninsky and T. J. White. Academic Press Inc., CA, USA, 1990. pp. xviii + 482. $39.95. ISBN 0-12-372181-40 , 1991 .

[21]  N. Sonenberg,et al.  High-level synthesis in Escherichia coli of functional cap-binding eukaryotic initiation factor eIF-4E and affinity purification using a simplified cap-analog resin. , 1988, Gene.

[22]  C. Marshall,et al.  MAP kinase kinase kinase, MAP kinase kinase and MAP kinase. , 1994, Current opinion in genetics & development.

[23]  Jiahuai Han,et al.  Independent human MAP-kinase signal transduction pathways defined by MEK and MKK isoforms , 1995, Science.

[24]  Jiahuai Han,et al.  Pro-inflammatory Cytokines and Environmental Stress Cause p38 Mitogen-activated Protein Kinase Activation by Dual Phosphorylation on Tyrosine and Threonine (*) , 1995, The Journal of Biological Chemistry.

[25]  John C. Lee,et al.  Hemopoietic Growth Factors with the Exception of Interleukin-4 Activate the p38 Mitogen-activated Protein Kinase Pathway* , 1997, The Journal of Biological Chemistry.

[26]  P. Cohen,et al.  Participation of a stress-activated protein kinase cascade in the activation of tyrosine hydroxylase in chromaffin cells. , 1997, European journal of biochemistry.

[27]  M. Gaestel,et al.  CREB is activated by UVC through a p38/HOG‐1‐dependent protein kinase , 1997, The EMBO journal.

[28]  M. Gaestel,et al.  3pK, a novel mitogen-activated protein (MAP) kinase-activated protein kinase, is targeted by three MAP kinase pathways , 1996, Molecular and cellular biology.

[29]  A. Ullrich,et al.  ERK6, a mitogen-activated protein kinase involved in C2C12 myoblast differentiation. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[30]  M. Cobb,et al.  Mitogen-activated protein kinase pathways. , 1997, Current opinion in cell biology.

[31]  D. Zechner,et al.  A Role for the p38 Mitogen-activated Protein Kinase Pathway in Myocardial Cell Growth, Sarcomeric Organization, and Cardiac-specific Gene Expression , 1997, The Journal of cell biology.

[32]  R. Erikson,et al.  Activation of protein serine/threonine kinases p42, p63, and p87 in Rous sarcoma virus-transformed cells: signal transduction/transformation-dependent MBP kinases. , 1992, Molecular biology of the cell.

[33]  J. Blenis,et al.  Signal transduction via the MAP kinases: proceed at your own RSK. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[34]  Jiahuai Han,et al.  The primary structure of p38 gamma: a new member of p38 group of MAP kinases. , 1996, Biochemical and biophysical research communications.

[35]  C. Widmann,et al.  MEKKs, GCKs, MLKs, PAKs, TAKs, and tpls: upstream regulators of the c-Jun amino-terminal kinases? , 1997, Current opinion in genetics & development.

[36]  M. Gaestel,et al.  Stress‐ and mitogen‐induced phosphorylation of the small heat shock protein Hsp25 by MAPKAP kinase 2 is not essential for chaperone properties and cellular thermoresistance. , 1994, The EMBO journal.

[37]  Tsonwin Hai,et al.  Tissue-specific Pattern of Stress Kinase Activation in Ischemic/Reperfused Heart and Kidney* , 1997, The Journal of Biological Chemistry.

[38]  C. Huang,et al.  The primary structure of a human MAP kinase activated protein kinase 2. , 1994, Biochemical and biophysical research communications.

[39]  J. Blenis,et al.  A Xenopus ribosomal protein S6 kinase has two apparent kinase domains that are each similar to distinct protein kinases. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[40]  D. Jackson,et al.  Visualization of replication factories attached to a nucleoskeleton , 1993, Cell.

[41]  A. Lin,et al.  Identification of c-Jun NH2-terminal Protein Kinase (JNK)-activating Kinase 2 as an Activator of JNK but Not p38* , 1997, The Journal of Biological Chemistry.

[42]  Philip R. Cohen,et al.  Activation of the novel stress‐activated protein kinase SAPK4 by cytokines and cellular stresses is mediated by SKK3 (MKK6); comparison of its substrate specificity with that of other SAP kinases , 1997, The EMBO journal.

[43]  L. Mahadevan,et al.  Parallel signal processing among mammalian MAPKs. , 1995, Trends in biochemical sciences.

[44]  T. Hunter,et al.  Phosphopeptide mapping and phosphoamino acid analysis by two-dimensional separation on thin-layer cellulose plates. , 1991, Methods in enzymology.

[45]  R. Treisman,et al.  The p38 and ERK MAP kinase pathways cooperate to activate Ternary Complex Factors and c‐fos transcription in response to UV light. , 1996, The EMBO journal.

[46]  L Bibbs,et al.  A MAP kinase targeted by endotoxin and hyperosmolarity in mammalian cells. , 1994, Science.

[47]  R. Treisman,et al.  The SRF accessory protein Elk-1 contains a growth factor-regulated transcriptional activation domain , 1993, Cell.

[48]  S. Noselli,et al.  MKK7 Is A Stress-activated Mitogen-activated Protein Kinase Kinase Functionally Related to hemipterous * , 1997, The Journal of Biological Chemistry.

[49]  Michael E. Greenberg,et al.  Opposing Effects of ERK and JNK-p38 MAP Kinases on Apoptosis , 1995, Science.

[50]  Jiahuai Han,et al.  Activation of the transcription factor MEF2C by the MAP kinase p38 in inflammation , 1997, Nature.

[51]  Xiaozhong Wang,et al.  Stress-Induced Phosphorylation and Activation of the Transcription Factor CHOP (GADD153) by p38 MAP Kinase , 1996, Science.

[52]  A. Guimond,et al.  Characterization of 45-kDa/54-kDa HSP27 kinase, a stress-sensitive kinase which may activate the phosphorylation-dependent protective function of mammalian 27-kDa heat-shock protein HSP27. , 1995, European journal of biochemistry.

[53]  Jiahuai Han,et al.  Characterization of the Structure and Function of a Novel MAP Kinase Kinase (MKK6) (*) , 1996, The Journal of Biological Chemistry.

[54]  A. Ashworth,et al.  Stimulation of the stress-activated mitogen-activated protein kinase subfamilies in perfused heart. p38/RK mitogen-activated protein kinases and c-Jun N-terminal kinases are activated by ischemia/reperfusion. , 1996, Circulation research.

[55]  M. Karin,et al.  Mitogen-activated protein kinase cascades and regulation of gene expression. , 1996, Current opinion in immunology.

[56]  T. Perring,et al.  Detection of plant virus coat proteins on whole leaf blots. , 1991, Analytical biochemistry.

[57]  J. Landry,et al.  Oxidative stress-induced actin reorganization mediated by the p38 mitogen-activated protein kinase/heat shock protein 27 pathway in vascular endothelial cells. , 1997, Circulation research.

[58]  John C. Lee,et al.  Identification of Mitogen-activated Protein (MAP) Kinase-activated Protein Kinase-3, a Novel Substrate of CSBP p38 MAP Kinase (*) , 1996, The Journal of Biological Chemistry.

[59]  M. Karin,et al.  JNK1: A protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain , 1994, Cell.

[60]  M. Su,et al.  Role of p38 and JNK mitogen-activated protein kinases in the activation of ternary complex factors , 1997, Molecular and cellular biology.

[61]  M. Goedert,et al.  SAP kinase‐3, a new member of the family of mammalian stress‐activated protein kinases , 1996, FEBS letters.

[62]  P. Cohen,et al.  A stress-activated kinase cascade can mediate the activation of tyrosine hydroxylase in chromaffin cells. , 1997, Biochemical Society transactions.

[63]  Michel Morange,et al.  A novel kinase cascade triggered by stress and heat shock that stimulates MAPKAP kinase-2 and phosphorylation of the small heat shock proteins , 1994, Cell.

[64]  P. Cohen,et al.  MAPKAP kinase‐2; a novel protein kinase activated by mitogen‐activated protein kinase. , 1992, The EMBO journal.

[65]  A. Sharrocks,et al.  Integration of MAP kinase signal transduction pathways at the serum response element. , 1995, Science.

[66]  C. Manthey,et al.  Molecular Cloning and Characterization of a Novel p38 Mitogen-activated Protein Kinase* , 1997, The Journal of Biological Chemistry.

[67]  B. Beutler,et al.  Interactive effects of the tumor necrosis factor promoter and 3'-untranslated regions. , 1991, Journal of immunology.

[68]  Jerry L. Adams,et al.  A protein kinase involved in the regulation of inflammatory cytokine biosynthesis , 1994, Nature.

[69]  Tony Hunter,et al.  MNK1, a new MAP kinase‐activated protein kinase, isolated by a novel expression screening method for identifying protein kinase substrates , 1997, The EMBO journal.

[70]  T. Hunter,et al.  Convergence of MAP kinase pathways on the ternary complex factor Sap‐1a , 1997, The EMBO journal.

[71]  John G. McWhirter,et al.  Parallel signal processing , 1989 .

[72]  Wei Guo,et al.  Characterization of the Structure and Function of a New Mitogen-activated Protein Kinase (p38β)* , 1996, The Journal of Biological Chemistry.

[73]  D. Moller,et al.  Regulation and Interaction of pp90rsk Isoforms with Mitogen-activated Protein Kinases* , 1996, The Journal of Biological Chemistry.

[74]  G. Nemerow,et al.  Apoptosis signaling pathway in T cells is composed of ICE/Ced-3 family proteases and MAP kinase kinase 6b. , 1997, Immunity.