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PMID: 16314523 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of NDR protein kinase by hydrophobic motif phosphorylation mediated by the mammalian Ste20-like kinase MST3.

Molecular and cellular biology ·Vol. 25 ·No. 24 ·2005-12-00 ·Pages 11019-29

Stegert MR, Hergovich A, Tamaskovic R, Bichsel SJ, Hemmings BA

Abstract

NDR protein kinases are involved in the regulation of cell cycle progression and morphology. NDR1/NDR2 protein kinase is activated by phosphorylation on the activation loop phosphorylation site Ser281/Ser282 and the hydrophobic motif phosphorylation site Thr444/Thr442. Autophosphorylation of NDR is responsible for phosphorylation on Ser281/Ser282, whereas Thr444/Thr442 is targeted by an upstream kinase. Here we show that MST3, a mammalian Ste20-like protein kinase, is able to phosphorylate NDR protein kinase at Thr444/Thr442. In vitro, MST3 selectively phosphorylated Thr442 of NDR2, resulting in a 10-fold stimulation of NDR activity. MOB1A (Mps one binder 1A) protein further increased the activity, leading to a fully active kinase. In vivo, Thr442 phosphorylation after okadaic acid stimulation was potently inhibited by MST3KR, a kinase-dead mutant of MST3. Knockdown of MST3 using short hairpin constructs abolished Thr442 hydrophobic motif phosphorylation of NDR in HEK293F cells. We conclude that activation of NDR is a multistep process involving phosphorylation of the hydrophobic motif site Thr444/2 by MST3, autophosphorylation of Ser281/2, and binding of MOB1A.

MeSH Terms
Adaptor Proteins, Signal Transducing Amino Acid Motifs Animals COS Cells Carrier Proteins/metabolism Catalytic Domain Chlorocebus aethiops Cytoplasm/chemistry Enzyme Activation Humans Hydrophobic and Hydrophilic Interactions Intracellular Signaling Peptides and Proteins Okadaic Acid/pharmacology Phosphorylation Protein Serine-Threonine Kinases/analysis,drug effects,genetics,metabolism Threonine/metabolism
Chemicals
Adaptor Proteins, Signal Transducing Carrier Proteins Intracellular Signaling Peptides and Proteins MOB1B protein, human Okadaic Acid Threonine STK24 protein, human MAP4K4 protein, human Protein Serine-Threonine Kinases STK38L protein, human
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Stegert Mario R
Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
Hergovich Alexander
Tamaskovic Rastislav
Bichsel Samuel J
Hemmings Brian A
References (52)
52 references, click to expand
  1. Ndr protein kinase is regulated by phosphorylation on two conserved sequence motifs.
    J Biol Chem. 1999 Nov 26;274(48):33847-50 PMID: 10567341
  2. Protein kinase Cdc15 activates the Dbf2-Mob1 kinase complex.
    Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7325-30 PMID: 11404483
  3. The tricornered gene, which is required for the integrity of epidermal cell extensions, encodes the Drosophila nuclear DBF2-related kinase.
    Genetics. 2000 Dec;156(4):1817-28 PMID: 11102376
  4. Both phosphorylation and caspase-mediated cleavage contribute to regulation of the Ste20-like protein kinase Mst1 during CD95/Fas-induced apoptosis.
    J Biol Chem. 2001 May 4;276(18):14909-15 PMID: 11278782
  5. MST1-JNK promotes apoptosis via caspase-dependent and independent pathways.
    Genes Cells. 2001 Jun;6(6):519-30 PMID: 11442632
  6. Caspase cleavage of MST1 promotes nuclear translocation and chromatin condensation.
    Proc Natl Acad Sci U S A. 2001 Aug 28;98(18):10148-53 PMID: 11517310
  7. MST4, a new Ste20-related kinase that mediates cell growth and transformation via modulating ERK pathway.
    Oncogene. 2001 Oct 4;20(45):6559-69 PMID: 11641781
  8. Yeast Cbk1 and Mob2 activate daughter-specific genetic programs to induce asymmetric cell fates.
    Cell. 2001 Dec 14;107(6):739-50 PMID: 11747810
  9. The IAP proteins: caspase inhibitors and beyond.
    Sci STKE. 2000 Aug 8;2000(44):pe1 PMID: 11752601
  10. Cloning of MASK, a novel member of the mammalian germinal center kinase III subfamily, with apoptosis-inducing properties.
    J Biol Chem. 2002 Feb 22;277(8):5929-39 PMID: 11741893
  11. Phosphorylation and dimerization regulate nucleocytoplasmic shuttling of mammalian STE20-like kinase (MST).
    J Biol Chem. 2002 Apr 5;277(14):12351-8 PMID: 11805089
  12. Caspase-catalyzed cleavage and activation of Mst1 correlates with eosinophil but not neutrophil apoptosis.
    Blood. 2002 May 1;99(9):3432-8 PMID: 11964314
  13. Determination of the affinity of each component of a composite quaternary transition-state analogue complex of creatine kinase.
    Biochemistry. 2002 Jun 4;41(22):6995-7000 PMID: 12033932
  14. The Saccharomyces cerevisiae Mob2p-Cbk1p kinase complex promotes polarized growth and acts with the mitotic exit network to facilitate daughter cell-specific localization of Ace2p transcription factor.
    J Cell Biol. 2002 Sep 2;158(5):885-900 PMID: 12196508
  15. Caspase activation of mammalian sterile 20-like kinase 3 (Mst3). Nuclear translocation and induction of apoptosis.
    J Biol Chem. 2002 Sep 13;277(37):34367-74 PMID: 12107159
  16. Crystal structure of an activated Akt/protein kinase B ternary complex with GSK3-peptide and AMP-PNP.
    Nat Struct Biol. 2002 Dec;9(12):940-4 PMID: 12434148
  17. The protein kinase complement of the human genome.
    Science. 2002 Dec 6;298(5600):1912-34 PMID: 12471243
  18. Mechanism of Ca2+-mediated regulation of NDR protein kinase through autophosphorylation and phosphorylation by an upstream kinase.
    J Biol Chem. 2003 Feb 28;278(9):6710-8 PMID: 12493777
  19. Regulation of mammalian STE20-like kinase 2 (MST2) by protein phosphorylation/dephosphorylation and proteolysis.
    J Biol Chem. 2003 Apr 4;278(14):11760-7 PMID: 12554736
  20. Specific inhibition of gene expression using a stably integrated, inducible small-interfering-RNA vector.
    EMBO Rep. 2003 Jun;4(6):609-15 PMID: 12776180
  21. NDR family of AGC kinases--essential regulators of the cell cycle and morphogenesis.
    FEBS Lett. 2003 Jul 3;546(1):73-80 PMID: 12829239
  22. hippo encodes a Ste-20 family protein kinase that restricts cell proliferation and promotes apoptosis in conjunction with salvador and warts.
    Cell. 2003 Aug 22;114(4):445-56 PMID: 12941273
  23. The Drosophila Mst ortholog, hippo, restricts growth and cell proliferation and promotes apoptosis.
    Cell. 2003 Aug 22;114(4):457-67 PMID: 12941274
  24. RAM: a conserved signaling network that regulates Ace2p transcriptional activity and polarized morphogenesis.
    Mol Biol Cell. 2003 Sep;14(9):3782-803 PMID: 12972564
  25. Hippo promotes proliferation arrest and apoptosis in the Salvador/Warts pathway.
    Nat Cell Biol. 2003 Oct;5(10):914-20 PMID: 14502294
  26. The Salvador partner Hippo promotes apoptosis and cell-cycle exit in Drosophila.
    Nat Cell Biol. 2003 Oct;5(10):921-7 PMID: 14502295
  27. The Drosophila Ste20 family kinase dMST functions as a tumor suppressor by restricting cell proliferation and promoting apoptosis.
    Genes Dev. 2003 Oct 15;17(20):2514-9 PMID: 14561774
  28. The inhibitors of apoptosis: there is more to life than Bcl2.
    Oncogene. 2003 Nov 24;22(53):8568-80 PMID: 14634619
  29. Survivin, versatile modulation of cell division and apoptosis in cancer.
    Oncogene. 2003 Nov 24;22(53):8581-9 PMID: 14634620
  30. YSK1 is activated by the Golgi matrix protein GM130 and plays a role in cell migration through its substrate 14-3-3zeta.
    J Cell Biol. 2004 Mar 29;164(7):1009-20 PMID: 15037601
  31. Initiation of cytokinesis is controlled through multiple modes of regulation of the Sid2p-Mob1p kinase complex.
    Mol Cell Biol. 2004 Apr;24(8):3262-76 PMID: 15060149
  32. Caspase activation - stepping on the gas or releasing the brakes? Lessons from humans and flies.
    Oncogene. 2004 Apr 12;23(16):2774-84 PMID: 15077141
  33. Regulation of NDR2 protein kinase by multi-site phosphorylation and the S100B calcium-binding protein.
    J Biol Chem. 2004 May 28;279(22):23806-12 PMID: 15037617
  34. Mammalian Sterile20-like kinase 1 and the regulation of apoptosis.
    Biochem Soc Trans. 2004 Jun;32(Pt3):485-8 PMID: 15157167
  35. Human Mob proteins regulate the NDR1 and NDR2 serine-threonine kinases.
    J Biol Chem. 2004 Jun 4;279(23):24444-51 PMID: 15067004
  36. Regulation of the MST1 kinase by autophosphorylation, by the growth inhibitory proteins, RASSF1 and NORE1, and by Ras.
    Biochem J. 2004 Jul 15;381(Pt 2):453-62 PMID: 15109305
  37. Identification and characterization of the nuclear import and export signals of the mammalian Ste20-like protein kinase 3.
    FEBS Lett. 2004 Aug 13;572(1-3):41-5 PMID: 15304321
  38. Mechanism of activation of NDR (nuclear Dbf2-related) protein kinase by the hMOB1 protein.
    J Biol Chem. 2004 Aug 20;279(34):35228-35 PMID: 15197186
  39. Mechanosensory neurite termination and tiling depend on SAX-2 and the SAX-1 kinase.
    Neuron. 2004 Oct 14;44(2):239-49 PMID: 15473964
  40. Control of dendritic branching and tiling by the Tricornered-kinase/Furry signaling pathway in Drosophila sensory neurons.
    Cell. 2004 Oct 15;119(2):245-56 PMID: 15479641
  41. Molecular cloning and characterization of a conserved nuclear serine(threonine) protein kinase.
    Proc Natl Acad Sci U S A. 1995 May 23;92(11):5022-6 PMID: 7761441
  42. Activation of the Ste20-like oxidant stress response kinase-1 during the initial stages of chemical anoxia-induced necrotic cell death. Requirement for dual inputs of oxidant stress and increased cytosolic [Ca2+].
    J Biol Chem. 1997 Nov 14;272(46):29372-9 PMID: 9361019
  43. The crystal structure of a phosphorylase kinase peptide substrate complex: kinase substrate recognition.
    EMBO J. 1997 Nov 17;16(22):6646-58 PMID: 9362479
  44. Fission yeast orb6, a ser/thr protein kinase related to mammalian rho kinase and myotonic dystrophy kinase, is required for maintenance of cell polarity and coordinates cell morphogenesis with the cell cycle.
    Proc Natl Acad Sci U S A. 1998 Jun 23;95(13):7526-31 PMID: 9636183
  45. Calcium regulation of Ndr protein kinase mediated by S100 calcium-binding proteins.
    EMBO J. 1998 Oct 15;17(20):5913-22 PMID: 9774336
  46. A constitutively active and nuclear form of the MAP kinase ERK2 is sufficient for neurite outgrowth and cell transformation.
    Curr Biol. 1998 Oct 22;8(21):1141-50 PMID: 9799732
  47. New methodologies for measuring protein interactions in vivo and in vitro.
    Curr Opin Struct Biol. 2005 Feb;15(1):4-14 PMID: 15718127
  48. Control of cell proliferation and apoptosis by mob as tumor suppressor, mats.
    Cell. 2005 Mar 11;120(5):675-85 PMID: 15766530
  49. The Ste20-like kinase Mst2 activates the human large tumor suppressor kinase Lats1.
    Oncogene. 2005 Mar 17;24(12):2076-86 PMID: 15688006
  50. Human NDR kinases are rapidly activated by MOB proteins through recruitment to the plasma membrane and phosphorylation.
    Mol Cell Biol. 2005 Sep;25(18):8259-72 PMID: 16135814
  51. MST, a physiological caspase substrate, highly sensitizes apoptosis both upstream and downstream of caspase activation.
    J Biol Chem. 2001 Jun 1;276(22):19276-85 PMID: 11278283
  52. Neuronal cell shape and neurite initiation are regulated by the Ndr kinase SAX-1, a member of the Orb6/COT-1/warts serine/threonine kinase family.
    Mol Biol Cell. 2000 Sep;11(9):3177-90 PMID: 10982409
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.24.11019-11029.2005
Published
2005-12-00
Pages
11019-29
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1316964
Subset
IM
External Links
PubMed source DOI: 10.1128/MCB.25.24.11019-11029.2005 PMC Full Text View journal details
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