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



1.Research Institution Yokoahama City University
 
2.Research Area Life Science
 
3.Research Field Cellular Signaling
 
4.Term of Project FY1996〜FY2000
 
5.Project Number 96L00305
 
6.Title of Project Regulation of Protein Kinases in Signal Transduction

7.Projetct Leader
Name Institution,Department Title of Position
Shigeo Ohno Yokohama City University School of Medicine Professor

8.Core Members

Names Institution,Department Title of Position
Syu-ichi Hirai Yokohama City University School of Medicine Associated Professor
Atsushi Suzuki Yokohama City University School of Medicine Lecturer
Keiko Mizuno Yokohama City University School of Medicine Assistant Professor

9.Cooperating Researchers

Names Institution,Department Title of Position
Tetsuo Ohnishi Yokohama City University School of Medicine Research Fellow
Yuki Sugiyama Yokohama City University School of Medicine Research Fellow
Masa-aki Nakaya Yokohama City University School of Medicine Research Fellow

10.Summary of Research Results

We focused several protein kinases that include protein kinaseC(PKC) delta and lambda, mixed lineage kinases, and DMPK, and analyzed their regulatory mechanism through identification of their binding proteins.
We found a novel member of the small heat shock protein family, MKBP, as a DMPK-specific binding protein. DMPK binds and stabilized DMPK, implying the role of DMPK as one of the components of a muscle-specific regulatory system that protoects muscle fibers from heat stress.
As for PKC, we found and identified ASIP, atypical PKC-specific interacting protein, as a physiological binding partner of atypical PKJKC isotypes. ASIP is a PDZ domain-containing adaptor-like protein and is the mammalian orthologue of C. elegans PAR-3 and Drosophila Bazooka, both of which are involved in the establishment of cell polarity. ASIP as well as aPKC localize to the tight junctions of vertebrate epithelial cells and are required for the establishment of epithelial cell polarith that include the development of epithelia-specific junctional structures.
We also found a novel member of the phosphatidylinositol kinase-related protein kinase (PIKK) family. This kinase was termed as hSMG-1 based on its close similarity to C. elegans SMG-1, one of the 7 smg genes involved in nonsense-mediated mRNA decay (NMD). In fact, we have succeeded in provide evidence supporting that hSMG-1 is involved in mammian NMD through phosphorylation of another component of the NMD system, hUpf1/SMG-2.

11.Key Words

(1)PKC、(2)DMPK、(3)small HSP
(4)stress response、(5)polarity、(6)epithela
(7)PAR、(8)aPKC、(9)cell-cell junctions

12.References

[Reference Articles]
Author Title of Article
Takaki, Y., Hirai, S., Manabe, N., Izumi, Y., Hirose, T., Nakaya, M., Suzuki, A., Mizuno, K., Akimoto, K., Shuin, T., and Ohno, S. Dynamic changes in protein components of the tight junction during liver regeneration
Journal Volume Year Pages Concerned
Cell & Tissue Research   in press  

Author Title of Article
Baba, M., Hirai, S., Kawakami, S., Kishida, T., Sakai, N., Kaneko, S., Yao, M., Shuin, T., Hosaka, M., and Ohno, S. Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth.
Journal Volume Year Pages Concerned
Oncogene   in press  

Author Title of Article
Noda, Y., Takeya, R., Ohno, S., Naito, S., Ito, T., and Sumimoto, H. Human homologues ofo the Caenorhabditis elegans cell polarity protein PAR6 as an adaptor that links the small GTPases Rac and Cdc42 to atypical protein kinase C.
Journal Volume Year Pages Concerned
Genes to Cells 6 2001 107-119

Author Title of Article
Suzuki, A., Yamanaka, T., Hashiba, K., Hirose, T., Izumi, Y., Ohnishi, T. and Ohno, S. Atypical PKC is involved in the establishment of epithelial polarity in mammalian cells.
Journal Volume Year Pages Concerned
J. Cell Biol. 152 2001 1183-1196

Author Title of Article
Nakaya, M., Fukui, A., Izumi, Y., Akimoto, K., Asashima, M., and Ohno, S. Meiotic Maturation Induces Animal-Vegetal Asymmetric Distribution of aPKC and ASIP/PAR-3 in Xenopus oocytes.
Journal Volume Year Pages Concerned
Development 127 2000 5021-5031

Author Title of Article
Kotani K, Ogawa W, Hashiramoto M, Onishi T, Ohno S., Kasuga M. Inhibition of insulin-induced glucose uptake by atypical protein kinase C isotype-specific interacting protein in 3T3-L1 adipocytes.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275 2000 26390-26395

Author Title of Article
Fukuyama, K., Yoshida, M., Yamashita, A., Deyama. T., Baba, M., Suzuki, A., Mohri, H., Ikezawa. Z., Nakajima. H., Hirai, S-I. and Ohno, S. MAPK upstream kinase(MUK)-binding inhibitory protein, a negative regulator of MUK/dual leucine zipper-bearing kinase/leucine zipper protein kinase.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275 2000 21247-21254

Author Title of Article
Sugiyama, Y., Suzuki, A., Kishikawa, M., Akutsu, R., Hirose, T., Waye, M. M. Y., Tsui, S. K. W., Yoshida, S. and Ohno, S. Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation.
Journal Volume Year Pages Concerned
J. Biol. Chem. 275 2000 1095-1104

Author Title of Article
Zhuang, S., Hisai, S. and Ohno, S. Hyperosmolality induces activation of cPKC and nPKC, a requirement for ERK1/2 activation in NIH/3T3 cells.
Journal Volume Year Pages Concerned
Am. J. Physiol. Cell Physiol. 278 2000 102-109

Author Title of Article
Okuda, H., Hirai, S., Takaki, Y., Kamata, M., Baba, M., Sakai, N., Kishida, T., Kaneko, S., Yao, M., Ohno, S. and Shuin, T. Direct interaction of the b-domain of VHL tumor suppressor protein with the regulatory domain of atypical PKC isotypes.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 263 1999 491-497

Author Title of Article
Koike, H., Tomioka, S., Sorimachi, H., Saido, T. C., Maruyama, K., Okuyama, A., Fujisawa-Sehara, A., Ohno, S., Suzuki, K. and Ishiura, S. MDC9 has an alpha-secretase activity responsible for processing the amyloid precursor protein.
Journal Volume Year Pages Concerned
Biochem. J. 343 1999 371-375

Author Title of Article
Imafuku, I., Masaki, T., Waragai, M., Takeuchi, S., Kawabata, M., Hirai, S., Ohno, S., Nee, L. E., Lippa, C. F., Kanazawa, I., Imagawa, M. and Okazawa, H. Presenilin 1 Suppresses the function of c-Jun homodimers via interaction with QM/Jif-1.
Journal Volume Year Pages Concerned
J. Cell Biol. 147 1999 121-133

Author Title of Article
Desai, D. S., Hirai, S., Kames, W. E. Jr., Niles, R. M. and Ohno, S. Cloning and characterization of the murine PKCa promoter: identification of a retinoic acid response element.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 263 1999 25-34

Author Title of Article
Miranti, C. K., Ohno, S. and Brugge, J. S. Protein kinase C regulates integrin-induced activation of the extracellular regulated kinase pathway upstream of Shc.
Journal Volume Year Pages Concerned
J. Biol. Chem. 274 1999 10571-10581

Author Title of Article
Izumi, Y., Hirata, M., Hasuwa, H., Iwamoto, R., Umata, T., Miyado, K., Tamai, Y., Kurisaki, T., Sehara-Fujisawa, A., Ohno, S. and Mekada, E. A metalloprotease-disintegrin, MDC9/Meltrin-g/ADAM9, and PKCd are involved in TPA-induced ectodomain shedding of membrane-anchored heparin-binding EGF-like growth factor.
Journal Volume Year Pages Concerned
EMBO J . 24 1998 7260-7272

Author Title of Article
Kotani, K., Ogawa, W., Matsumoto, M., Kitamura, T., Sakaue, H., Hino, Y., Miyake, K., Sano, W., Akimoto, K., Ohno, S. and Kasuga M.: Requirement of atypical PKC1 for insulin stimulation of glucose uptake but not for Akt activation in 3T3-L1 adipocytes.
Journal Volume Year Pages Concerned
Molecular and Cellular Biol. 18 1998 6971-6982

Author Title of Article
Izumi, Y., Hirose, T., Tamai, Y., Hirai, S-I., Nagashima, Y., Fujimoto, T., Tabuse, Y., Kemphues, K. J. and Ohno, S. An atypical PKC directly associates and co-localizes at the epithelial tight junction with ASIP, a mammalian homologue of Caenorhabditis elegans polarity protein PAR-3.
Journal Volume Year Pages Concerned
J. Cell Biol. 143 1998 95-106

Author Title of Article
Akimoto, K., Nakaya, M., Yamanaka, T., Tanaka, J., Matsuda, S., Weng, Q-P., Avruch, J. and Ohno, S. Atypical PKC1 binds and regulates p70 S6 kinase.
Journal Volume Year Pages Concerned
Biochem. J. 335 1998 417-424

Author Title of Article
Tabuse, Y., Izumi, Y., Piano, F., Kemphues, K. J., Miwa, J. and Ohno, S. Atypical Protein Kinase C Cooperates with PAR-3 to Establish Embryonic Polarity in Caenorhabditis elegans.
Journal Volume Year Pages Concerned
Development 125 1998 3607-3614

Author Title of Article
Yuasa, T., Ohno, S., Kehrl, J. H. and Kyriakis, J. M. Tumor necrosis factor signaling to stress-activated protein kinase(SAPK)/Jun NH2-terminal kinase(JNK)and p38. Germinal center kinase couples TRAF2 to mitogen-activated protein kinase/ERK kinase kinase 1 and SAPK while receptor interacting protein associates with a mitogen-activated protein kinase kinase kinase upstream of MKK6 and p38.
Journal Volume Year Pages Concerned
J. Biol. Chem. 273 1998 22681-22692

Author Title of Article
Nyui, N., Tamura, K., Mizuno, K., Ishigami, T., Kihara, M., Ochiai, H., Kimura, K., Umemura, S., Ohno, S., Taga, T. and Ishii, M. gp130 is involved in stretch-induced MAP kinase activation in cardiac myocytes.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 245 1998 928-932

Author Title of Article
Hirai, S-I., Noda, K., Moriguchi, T., Nishida, E., Yamashita, A., Deyama, T., Fukuyama, K. and Ohno, S. Differential activation of two JNK activators, MKK7 and SEK1, by MKN28-derived nonreceptor serine/threonine kinase/mixed lineage kinase 2.
Journal Volume Year Pages Concerned
J. Biol. Chem. 273 1998 7406-7412

Author Title of Article
Suzuki, A., Sugiyama, Y., Hayashi, Y., Nyu-i, N., Yoshida, M., Nonaka, I., Ishiura, S., Arahata, K. and Ohno, S. MKBP, a novel member of the small heat shock protein family, binds and activates the myotonic dystrophy protein kinase(DMPK).
Journal Volume Year Pages Concerned
J. Cell Biol. 140 1998 1113-1124

Author Title of Article
Zhuang, S., Hirai, S-I, Mizuno, K., Suzuki, A., Akimoto, K., Izumi, Y., Yamashita, A. and Ohno, S. Involvement of protein kinase C in the activation of extracelluar signal-regulated kinase 1/2 by UVC irradiation.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 240 1997 273-278

Author Title of Article
Ichimura, T., Ito, M., Itagaki, C., Takahashi, M., Horigome, T., Omata, S., Ohno, S. and Isobe, T. The 14-3-3 protein binds its target proteins with a common site located towards the C-terminus.
Journal Volume Year Pages Concerned
FEBS Letter 413 1997 273-276

Author Title of Article
Mizuno, K., Noda, K., Araki, T., Imaoka, T., Kobayashi, Y., Akita, Y., Shimonaka, M., Kishi, S. and Ohno, S. The proteolytic cleavage of protein kinase C isotypes, which generates kinase and regulatory fragments, correlates with Fas-mediated and 12-0-tetradecanoyl-phorbol-13-acetate-induced apotosis.
Journal Volume Year Pages Concerned
Eur. J., Biochem. 250 1997 7-18

Author Title of Article
Nyui, N, Tamura, K., Mizuno, K., Ishigami, T., Hibi, K., Yabana, M., Kihara, M., Fukamizu, A., Ochiai, H., Umemura, S., Murakami, K., Ohno, S. and Ishii, M. Stretch-induced map kinase activation in cardiomyocytes of angiotensinogen-deficientmice.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 233 1997 36-41

Author Title of Article
Osada, S., Izawa, M., Saito, R., Mizuno, K., Suzuki, A., Hirai, S. and Ohno. S. YSK1, a novel mammalian protein kinase structurally related to Ste20 and SPS1, but not involved in the known MAPK pathways.
Journal Volume Year Pages Concerned
Oncogene 14 1997 2047-2057

Author Title of Article
Lu, Z., Hornia, A., Jiang, Y-W., Frankel, P., Zang Q., Ohno, S. and Foster, D. A. Tumor-promotion by depleting cells of protein kinase Cd.
Journal Volume Year Pages Concerned
Molecular and Cellular Biol. 17 1997 3418-3428

Author Title of Article
Fukumoto, S., Nishizawa, Y., Hosoi, M., Koyama, H., Yamakawa, K., Ohno, S. and Morii, H. Protein kinase C delta isoform inhibits the proliferation of vascular smooth muscle cells through suppressiong G1 cyclins expression.
Journal Volume Year Pages Concerned
J. Biol. Chem. 272 1997 13816-13822

Author Title of Article
Hirai, S., Katoh, M., Terada, M., Kyriakis, J. M., Zon, L. I., Avruch, J. and Ohno, S. MST/MLK2, a member of the mixed lineage kinase family, directly phosphorylates and activates SEK1, an activator of JNK/SAPK.
Journal Volume Year Pages Concerned
J. Biol. Chem. 272 1997 15167-15173

Author Title of Article
Izumi, Y., Hirai, S., Tamai, Y., Fujise-Matsuoka, A., Nishimura, Y. and Ohno, S. A protein kinase C delta-binding protein SRBC whose expression is induced by serum starvation.
Journal Volume Year Pages Concerned
J. Biol. Chem. 272 1997 7381-7389

Author Title of Article
Osada, S., Izawa, M., Koyama, T., Hirai, S. and Ohno, S. A domain containing the Cdc42/Rac interactive binding(CRIB)region of p65PAK inhibits transcriptional activation and cell transformation mediated by the Ras-Rac pathway.
Journal Volume Year Pages Concerned
FEBS Letter 404 1997 227-233

Author Title of Article
Ohno, S. The distinct biological potential of PKC isotypes.
Journal Volume Year Pages Concerned
Protein Kinase C   1997 75-93

Author Title of Article
Chalfant, C. E., Ohno, S., Konno, Y., Fisher, A. A., Watson, J. E. and Cooper, D. R. A carboxy-terminal deletion mutant of PKCbeta II inhibits insulin-stimulated 2-deoxyglucose uptake in rat L6 skeletal muscle cells.
Journal Volume Year Pages Concerned
Molecular Endocrinology 10 1996 1273-1281

Author Title of Article
Ueda, Y., Hirai, S., Osada, S., Suzuki, A., Mizuno, K. and Ohno, S. Protein kinase C d activates the MEK-ERK pathway in a manner independent of Ras and dependent on Raf.
Journal Volume Year Pages Concerned
J. Biol. Chem. 271 1996 23512-23519

Author Title of Article
Rana, A., Gallo, K., Godowski, P., Hirai, S., Ohno, S., Zon, L., Kyriakis, J. M. and Auruch, J. The mixed lineage kinase SPRK phosphorylates and activates the stress-activated protein kinase activator, SEK-1.
Journal Volume Year Pages Concerned
J. Biol. Chem. 271 1996 19025-19028

Author Title of Article
Nakaigawa, N., Hirai, S.-I., Mizuno, K., Suzuki, A., Shuin, T., Hosaka, M. and Ohno, S. Differential effects of overexpression of PKC α and PKC δ/ε on cellular E2F activity at late G1 phase.
Journal Volume Year Pages Concerned
Biochem. Biophys. Res. Commun. 222 1996 93-100

Author Title of Article
Hirano, M., Osada, S., Aoki, T., Hirai, S., Hosaka, M., Inoue, J., and Ohno, S. MEK kinase is involved in TNFa-mediated activation of NFkB and degradation of IkBa.
Journal Volume Year Pages Concerned
J. Biol. Chem. 271 1996 13324-13238

Author Title of Article
Ueda, E., Ohno, S., Kuroki, T., Livneh, E., Yamada, K., Yamanishi, K., Yasuno, H. The h isoform of protein kinase C mediates transcriptional activation of the human transglutaminase 1 gene.
Journal Volume Year Pages Concerned
J. Biol. Chem. 271 1996 9790-9794

Author Title of Article
Akimoto, K., Takahashi, R., Moriya, S., Nishioka, N., Takayanagi, J., Kimura, K., Fukui, Y., Osada, S., Mizuno, K., Hirai, S., Kazlauskas, A. and Ohno, S. EGF and PDGF receptors activate atypical PKC1 through phosphatidylinositol 3-kinase.
Journal Volume Year Pages Concerned
EMBO J. 15 1996 788-798

Author Title of Article
Hirai, S., Izawa, N., Osada, S., Spyrou, G. and Ohno, S. Activation of the JNK pathway by distantly related protein kinases, MEKK and MUK.
Journal Volume Year Pages Concerned
Oncogene. 12 1996 641-630

Author Title of Article
Moriya, S., Kazlauskas, A., Akimoto, K., Hirai, S., Mizuno, K., Takenawa, T., Fukui, Y., Watanabe, Y., Ozaki, S. and Ohno, S. PDGF activates protein kinase Ce through redundant and independent signaling pathways involving phosphpholipase Cg or phosphatidylinositol 3-kinase.
Journal Volume Year Pages Concerned
Proc Natl. Acad. Sci. U. S. A. 93 1996 151-155


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