| 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 |
| 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 α-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 transduction、(2)Protein kinases、(3)PKN
(4)PKC、(5)Small GTP binding Protein、(6)Cytoskeletal protein
(7)Apoptosis、(8)Cell cycle、(9)Anchoring protein
12.References
| 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 α-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 γ-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 αB-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 β , a novel isoform of protein kinase PKN: expression and arachidonic acid dependency are different from those of PKN α. | |||
| 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 δ and protein kinase B α 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 β 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 α and protein kinase C on phosphorylation of tau protein by glycogen synthase kinase-3 β. | |||
| 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 δ 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 ε 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 ε isozyme of protein kinase C. | |||
| Journal | Volume | Year | Pages Concerned | |
| J. Biol. Chem. | in press. | |||