Summary of Research Project Results Under the JSPS FY2000
"Research for the future Program"



1.Research Institution@Kobe University
@
2.Research Area@Life Sciences
@
3.Research Field@Cellular Signaling
@
4.Term of Project@FY1996`FY2000
@
5.Project Number@96L00306
@
6.Title of Project@Signaling network by protein kinases

7.Projetct Leader
Name Institution,Department Title of Position
Ono, Yoshitaka. Kobe University, Biosignal Research Center Professor

8.Core Members

Names Institution,Department Title of Position
Mukai, Hideyuki Kobe University, Biosignal Research Center Associate Professor
Kuno, Takayoshi Kobe University, School of Medicine Professor
Sakamoto, Hiroshi Kobe University, Faculty of Science Professor

9.Cooperating Researchers

Names Institution,Department Title of Position
Takahashi, Mikiko Kobe University, Biosignal Research Center Assistant Professor
Miyamoto, Masaaki Kobe University, Graduate School of Science and Technology Associate Professor

10.Summary of Research Results

PKN in a serine/threonine protein kinase that has a catalytic domain highly homologous to PKC in the carboxyl-terminal region and a unique regulatory domain in the amino-terminal region. The amino-terminal region of PKN contains three repeats of a leucine zipper-like motif and its kinase activity is stimulated by fatty acids such as arachidonic acid. We found that PKN is one of the downstream targets of small GTPase Rho, that regulates actin-based cytoskeletal structures, including focal adhesions, stress fibers and the contractile ring. We also found that PKN associates with cytoskeletal proteins such as intermediate filament proteins and actin cross-linking protein -actinin, suggesting that PKN participates in cytoskeletal reorganization. Furthermore, we reported that PKN translocates from the cytosol to the nucleus of fibroblasts upon exposure to stress such as heat shock and serum starvation and that PKN is cleaved during apoptosis presumably by caspase-3, which generates a constitutively active kinase fragment. And we found that PKN directly phosphorylates and inhibits mitotic regulatory phosphatase Cdc25C. These results raise the possibility that PKN has important roles both in the cytoskeletal reorganization and nuclear events including regulation of gene expression and cell cylcle.

11.Key Words

(1)Signal transductionA(2)Protein kinasesA(3)PKN
(4)PKCA(5)Small GTP binding ProteinA(6)Cytoskeletal protein
(7)ApoptosisA(8)Cell cycleA(9)Anchoring protein

12.References

[Reference Articles]
Author Title of Article
Watanabe, G. Protein kinase N (PKN) and PKN-related protein Rhophilin as targets of small GTPase Rho.
Journal Volume Year Pages Concerned
Science 271 1996 645-648

Author Title of Article
Amano, M. Identification of a putative target for Rho as the serine-threonine kinase, PKN.
Journal Volume Year Pages Concerned
Science 271 1996 648-650

Author Title of Article
Mukai, H. PKN associates and phosphorylates the head-rod domaine of neurofilament protein.
Journal Volume Year Pages Concerned
J. Biol. Chem. 271 1996 9816-9822

Author Title of Article
Kitagawa, M. The role of the unique motifs in the amino-terminal region of PKN on its enzymatic activity.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 220 1996 963-968

Author Title of Article
Shibata, H. Characterization of the interaction between RhoA and the amino-terminal region of PKN.
Journal Volume Year Pages Concerned
FEBS Lett. 385 1996 221-224

Author Title of Article
Konishi, H. Activation of RAC-protein kinase by heat shock and hyperosmolarity stress through a pathway independent of phosphatidylinositol 3-kinase.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 93 1996 7639-7643

Author Title of Article
Mukai, H. Translocation of PKN from the cytosol to the nucleus induced by stresses.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 93 1996 10195-10199

Author Title of Article
Matsuzaki, M. Isolation of the active form of RAC-protein kinase (PKB/Akt) from transfected COS-7 cells treated with heat shock stress and effects of phosphatidylinositol 3, 4, 5- trisphosphate and phosphatidylinositol 4, 5-bisphosphate on its enzyme activity.
Journal Volume Year Pages Concerned
FEBS Lett. 396 1996 305-308

Author Title of Article
Mukai, H. Interaction of PKN with -actinin.
Journal Volume Year Pages Concerned
J. Biol. Chem. 272 1997 4740-4746

Author Title of Article
Konishi, H. Activation of protein kinase B (Akt/RAC-protein kinase) by cellular stress and its association with heat shock protein Hsp27.
Journal Volume Year Pages Concerned
FEBS Lett. 410 1997 493-498

Author Title of Article
Matsuzawa, K. Domain-specific phosphorylation of vimentin and glial fibrillary acidic protein by PKN.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 234 1997 621-625

Author Title of Article
Kitagawa, M. Molecular cloning and characterization of a novel mitochondrial phosphoprotein, MIPP65, from rat liver.
Journal Volume Year Pages Concerned
Exp. Cell Res. 235 1997 71-78

Author Title of Article
Konishi, H. Activation of protein kinase C by tyrosine phosphorylation in response to H2O2.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 94 1997 11233-11237

Author Title of Article
Sakai, N. Direct visualization of the translocation of the -subspecies of protein kinase C in living cells using fusion proteins with green fluorescent protein.
Journal Volume Year Pages Concerned
J. Cell Biol. 139 1997 1465-1476

Author Title of Article
Bartsch, J. W. Protein kinase N (PKN) gene (PRKCL1/Prkcl1) maps to human chromosome 19p12-p13.1 and mouse chromosome 8 with close linkage to the myodystrophy (myd) mutation.
Journal Volume Year Pages Concerned
Genomics 49 1998 129-132

Author Title of Article
Takanaga, H. PKN interacts with a paraneoplastic cerebellar degeneration-associated antigen, which is a potential transcription factor.
Journal Volume Year Pages Concerned
Exp. Cell Res. 241 1998 363-372

Author Title of Article
Tokunaga, C. Molecular cloning and characterization of a novel protein kinase C-interacting protein with structural motifs related to RBCC family proteins.
Journal Volume Year Pages Concerned
Biochem Biophys. Res. Commun. 244 1998 353-359

Author Title of Article
Standaert, M. Comparative effects of GTP gamma S and insulin on the activation of Rho, phosphatidylinositol 3-kinase, and protein kinase N in rat adipocytes-relationship to glucose transport.
Journal Volume Year Pages Concerned
J. Biol. Chem. 273 1998 7470-7477

Author Title of Article
Takahashi, M. Proteolytic activation of PKN by Caspase-3 or related protease during apoptosis.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 95 1998 11566-11571

Author Title of Article
Kawamata, T. A protein kinase, PKN, accumulates in Alzheimer neurofibrillary tangles and associated endoplasmic reticulum-derived vesicles and phosphorylates Tau protein.
Journal Volume Year Pages Concerned
J. Neurosci. 18 1998 7402-7410

Author Title of Article
Hashimoto, T. Localization of PKN mRNA in the rat brain.
Journal Volume Year Pages Concerned
Mol. Brain Res. 59 1998 143-153

Author Title of Article
Kitagawa, M. The role of PKN in the regulation of B-crystallin expression via heat shock transcription factor 1.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 252 1998 561-565

Author Title of Article
Tanaka, M. Identification of myosin II as a binding protein to the PH domain of protein kinase B.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 255 1999 169-174

Author Title of Article
Yamazaki, M. Interaction of the small G protein RhoA with the C terminus of human phospholipase D1.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 6035-6038

Author Title of Article
Bandyopadhyay, G. Effects of transiently expressed atypical (zeta, lambda), conventional (alpha, beta) and novel (delta, epsilon) protein kinase C isoforms on insulin-stimulated translocation of epitope-tagged GLUT4 glucose transporters in rat adipocytes: specific interchangeable effects of protein kinases C-zeta and C-lambda.
Journal Volume Year Pages Concerned
Biochemical J. 337 1999 461-470

Author Title of Article
Takahashi, M. Characterization of a novel giant scaffolding protein, CG-NAP, that anchors multiple signaling enzymes to centrosome and the Golgi apparatus.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 17267-17274

Author Title of Article
Yoshinaga, C. Mutational analysis of the regulatory mechanism of PKN: the regulatory region of PKN contains an arachidonic acid-sensitive autoinhibitory domain.
Journal Volume Year Pages Concerned
J. Biochem. 126 1996 475-484

Author Title of Article
Oishi, K. Identification and characterization of PKN , a novel isoform of protein kinase PKN: expression and arachidonic acid dependency are different from those of PKN .
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 261 1996 808-814

Author Title of Article
Banno, Y. Differential phospholipase D activation by bradykinin and sphingosine 1-phosphate in NIH 3T3 fibroblasts overexpressing gelsolin.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1996 27385-27391

Author Title of Article
Takaishi, H. Regulation of nulclear translocation of forkhead transcription factor AFX by protein kinase B.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 96 1999 11836-11841

Author Title of Article
Roy K. Unsaturated fatty acd-activated protein kinase (PKx) from goat testis cytosol.
Journal Volume Year Pages Concerned
Biochim. Biophys. Acta 1434 1999 161-169

Author Title of Article
Konishi, H. Opposing effects of protein kinase C and protein kinase B on H2O2-induced apoptosis in CHO cells.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 264 1999 840-846

Author Title of Article
Tan, Z. Effects of local anesthetics on phospholipase D activity in differentiated human promyelocytic leukemic HL60 cells.
Journal Volume Year Pages Concerned
Biochem. Pharmacol. 58 1999 1881-1889

Author Title of Article
Konishi, H. Andjelkovic, M., Hemmings, B. A., and Kikkawa, U. Activation of protein kinase B induced by H2O2 and heat shock through distinct mechanisms dependent and independent of phosphatidylinositol 3-kinase.
Journal Volume Year Pages Concerned
J. Biochem. 126 1999 1136-1143

Author Title of Article
Standaert, M. L. Effects of knockout of the protein kinase C gene on glucose transport and glucose homeostasis.
Journal Volume Year Pages Concerned
Endocrinology 140 1999 4470-4477

Author Title of Article
Shibata, H. Interaction of PKN with a neuron-specific basic helix-loop-helix transcription factor, NDRF/NeuroD2.
Journal Volume Year Pages Concerned
Mol. Brain Res. 74 1999 126-134

Author Title of Article
Dong, L. Q. Phosphorylation of protein kinase N by phosphoinositide-dependent protein kinase-1 mediates insulin signals to the actin cytoskeleton.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 97 2000 5089-5094

Author Title of Article
Nagumo, H. Rho kinase inhibitor HA-1077 prevents Rho-medicated myosin phosphatase inhibitation in smooth muscle cells.
Journal Volume Year Pages Concerned
Am. J. Physiol. Cell Physiol. 278 2000 57-65

Author Title of Article
Sumioka, K. Induction of a 55-kDa PKN cleavage product by ischemia/reperfusion model in the rat retina.
Journal Volume Year Pages Concerned
Inv. Ophth. Vis. Sci. 41 2000 29-35

Author Title of Article
Isagawa, T. Dual effects of PKN and protein kinase C on phosphorylation of tau protein by glycogen synthase kinase-3 .
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 273 2000 209-212

Author Title of Article
Yamamoto, T. H2O2-induced tyrosine phosphorylation of protein kinase C by a mechanism independent of inhibition of protein-tyrosine phosphatase in CHO and COS-7 cells.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 273 2000 960-966

Author Title of Article
Gao, Q. PKN binds and phosphorylates human Papillomavirus E6 oncoprotein.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275 2000 14824-14830

Author Title of Article
Takahashi, M. Association of immature hypo-phosphorylated protein kinase C with an anchoring protein CG-NAP.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275 2000 34592-34596

Author Title of Article
Misaki, K. PKN delays mitotic timing by inhibition of Cdc25C: Possible involvement of PKN in the regulation of cell division.
Journal Volume Year Pages Concerned
Proc. Natl. Acad. Sci. USA 98 2001 125-129

Author Title of Article
Taniguchi, T. Phosphorylation of tau is regulated by PKN.
Journal Volume Year Pages Concerned
J. Biol. Chem. @ @ in press.

Author Title of Article
Matsumoto, M. Inhibition of insulin-induced activation of Akt by a kinase-deficient mutant of the isozyme of protein kinase C.
Journal Volume Year Pages Concerned
J. Biol. Chem. @ @ in press.


back