2001 年度実績報告書

mRNAの品質管理システム(mRNAサーベイランス)の分子機序:ヒトSMG1の構造と機能の解析

研究課題

研究課題/領域番号	13480211
研究機関	横浜市立大学
研究代表者	大野茂男横浜市立大学, 医学部, 教授 (10142027)
研究分担者	秋本和憲横浜市立大学, 医学部, 助手 (70285104) 平井秀一横浜市立大学, 医学部, 助教授 (80228759)
キーワード	ナンセンス変異 / mRNA分解 / mRNAサーベイランス / p53 / プロテインキナーゼ / リン脂質キナーゼ / RNAヘリカーゼ / ATPアーゼ
研究概要	新規のリン脂質キナーゼ類似のプロテインキナーゼ(分子量400万以上の巨大タンパク質)を偶然に見いだしその性状解析を進めてきた。最近になり、これが線虫のRNA surveillanceの主役であるSMG1の、ヒト版であることを見いだした。本研究では、ヒトSMG1やそのキナーゼ欠損変異体などのcDNA発現系、阻害剤、グロビン遺伝子の変異体をレポーターとしたNMDアッセイ系、など既に我々が独自に準備した材料を武器として、ヒトSMG1が、実際にRNA surveillance機構に関わることを明かとした。さらに、ヒトSMG1の基質タンパク質として、hUPF1/SMG-2を同定すると同時に、そのhSMG-1によるリン酸化部位分子群を決定し、これが細胞内でも起きていることを証明した。さらに、hSMG-1が、hUPF1/SMG-2などを含むいわゆるサーベイランス複合体の構成員であることを明らかとした。また、精製hSMG-1を用いて、その阻害剤を検索し、PIKKファミリーの阻害剤であるwortmannninやcaffeinがhSMG-1のキナーゼ活性を細胞レベルでも阻害することを明らかとすると同時に、これらの阻害剤を用いて、細胞内在性の変異遺伝子のNMDを抑制できることを初めて明らかとした。これを用いることにより、変異遺伝子を発見する新しい手法の可能性を提示したことにもなる。さらに、線虫のSMG遺伝子群のほ乳類版を軒並み同定すると同時にその構造や分子間相互作用を明らかにした。これらの一連の実験を通じて、ほ乳類のNMDに関わる分子機構の更なる解析の道具立てがそろいつつある。

研究成果
(17件)

すべてその他

すべて文献書誌 (17件)

[文献書誌] Mishima, A., Suzuki, A., Enaka, M., Hirose, T, Mizuno, K., Ohnishi, T, Mohri, H., Ishigatsubo, Y., Ohno, S.: "Ectopic expression of PAR-3 disturbs the equilibrium between the polarized and non-polarized states of mammalian epithelial cells"Genes to Cells. (in press).
[文献書誌] Hirose, T, Izumi, Y., Nagashima, Y., Tamai-Nagai, Y., Kurihara, H., Sakai, T., Suzuki, Y., Yamanaka, T., Suzuki, A., Mizuno, K., Ohno, S.: "Involvement of ASIP/PAR-3 in the Promotion of Epithelial Tight Junction Formation"J. Cell Science. (in press).
[文献書誌] Miyamoto, A., Nakayama, K., Imaki, H., Hirose, S., Jiang, Y., Abe, M., Tsukiyama, T., Nagahama, H., Ohno, S., Hatakeyama, S., Nakayama, K-I.: "Increased proliferation of B cells and autoimmune disease in protein kinase C-delta-deficient mice"Nature. (in press).
[文献書誌] Hiwasa, T., Nakata, M., Nakata, M., Ohno, S., Maki M., Suzuki, K., Takiguchi, M.: "Regulation of Transformed State by Calpastatin via PKCepsilon in NIH3T3 Mouse Fibroblasts"Biochem. Biophys. Res. Commun.. 290. 510-517 (2002)
[文献書誌] Harbers, M., Nomura, T., Ohno, S., Ishii, S.: "Intracellular localization of the ret finger protein depends on a functional nuclear export signal and protein kinase C activation"J. Biol Chem.. 276. 48596-48607 (2001)
[文献書誌] Okuda, H., Saitoh, K., Hirai, Si., S., Iwai, K., Takak Y., Baba, M., Minato, N., Ohno, S., Shuin, T.: "The Von Hippel-Lindau (VHL) tumor suppressor protein mediates ubiquitination of activated atypical protein kinase C"J Biol Chem.. 276. 43611-43617 (2001)
[文献書誌] Ohno, S.: "Intercellularjunctions and cellular polarity : the PAR-aPKC complex, a conserved core cassette playing fundamental roles in cell polarity"Curr. Opin. Cell Biol.. 13. 641-648 (2001)
[文献書誌] Matstuda, S., Yasukawa, T., Homma, Y., Ito, Y., Niikura, T., Hiraki, T., Hirai, S., Ohno, S., Kita, Y., Kawasumi, M., Kouyama, K., Yamamoto, T., Kyriakis, J.M., Nishimoto, I.: "c-Jun n-terminal kinase (jnk)-interacting protein-1b/islet-brain-1 scaffolds alzheimer's amyloid precursorprotein with juk"J. Neurosci.. 21. 6597-6607 (2001)
[文献書誌] Zhao, X., Murata, T., Ohno, S., Day, N., Song, J., Nomura, N., Nakahara, T., Yokoyama, K.K.: "Protein kinase Ca plays a critical role in mannosylerythritol lipid-induced differentiation of melanoma B16 cells"J. Biol. Chem.. 276. 39903-39910 (2001)
[文献書誌] Yamashita, A., Ohnishi, T., Kashima, I., Taya, Y., Ohno, S.: "Human SMG-1, a novel phosphatidylinositol 3-kinase-related protein kinase, associates with components of the mRNA surveillance complex and is involved in the regulation of nonsense-mediated mRNA decay"Genes Dev.. 15. 2215-2228 (2001)
[文献書誌] Ebnet, K., Suzuki, A., Horikoshi, Y., Hirose, T., Meyer, Zu., Brickwedde, M.K., Ohno, S., Vestweber, D.: "The cell polarity protein ASIP/PAR-3 directly associates with junctional adhesion molecule (JAM)"EMBO. J.. 20. 3738-3748 (2001)
[文献書誌] Yamanaka, T., Horikoshi, Y., Suzuki, A., Sugiyama, Y., Kitamura, K., Maniwa, R., Nagai, Y., Yamashita, A., Hirose, T., Ishikawa, H., Ohno, S.: "PAR-6 regulates aPKC activity in a novel way and mediates cell-cell contact-induced formation of the epithelial junctional complex"Genes Cells. 6. 721-731 (2001)
[文献書誌] Yamaji, S., Suzuki, A., Sugiyama, Y., Koide, Y., Yoshida, M., Kanamori.H., Mohri, H., Ohno, S., Ishigatsubo, Y.: "Anovel integrin-linked kinase-binding protein, affixin, is involved in the early stage of cell-substrate interaction"J. Cell. Biol.. 153. 1251-1264 (2001)
[文献書誌] Takaki, Y., Hirai, S., Manabe, N., Izumi, Y., Hirose, T., Nakaya, M., Suzuki, A., Mizuno, K., Akimoto, K., Tsukita, S., Shuin, T., Ohno, S.: "Dynamic changes in protein components of the tight junction during liver regeneration"Cell Tissue Res.. 305. 399-409 (2001)
[文献書誌] Baba, M., Hirai, S., Kawakami, S., Kishida, T., Sakai, N., Kaneko, S., Yao, M., Shuin, T., Kubota, Y., Hosaka, M., Ohno, S.: "Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth"Oncogene. 20. 2727-2736 (2001)
[文献書誌] Noda, Y., Takeya, R., Ohno, S., Naito, S., Ito, T., Sumimoto, H.: "Human homologues of the Caenorhabditis elegans cell polarity protein PAR6 as an adaptor that links the small GTPases Rac and Cdc42 to atypical protein kinase C"Genes Cells. 6. 107-119 (2001)
[文献書誌] Suzuki, A., Yamanaka, T., Hirose, T., Manabe, N., Mizuno, K., Shimizu, M., Akimoto, K., Izumi, Y., Ohnishi, T., Ohno, S.: "Atypical protein kinase C is involved in the evolutionarily conserved par protein complex and plays a critical role in establishing epithelia-specific junctional structures"J. Cell Biol.. 152. 1183-1196 (2001)

2001 年度 実績報告書

mRNAの品質管理システム(mRNAサーベイランス)の分子機序:ヒトSMG1の構造と機能の解析

研究代表者

大野 茂男 横浜市立大学, 医学部, 教授 (10142027)

研究成果

[文献書誌] Mishima, A., Suzuki, A., Enaka, M., Hirose, T, Mizuno, K., Ohnishi, T, Mohri, H., Ishigatsubo, Y., Ohno, S.: "Ectopic expression of PAR-3 disturbs the equilibrium between the polarized and non-polarized states of mammalian epithelial cells"Genes to Cells. (in press).

[文献書誌] Hirose, T, Izumi, Y., Nagashima, Y., Tamai-Nagai, Y., Kurihara, H., Sakai, T., Suzuki, Y., Yamanaka, T., Suzuki, A., Mizuno, K., Ohno, S.: "Involvement of ASIP/PAR-3 in the Promotion of Epithelial Tight Junction Formation"J. Cell Science. (in press).

[文献書誌] Miyamoto, A., Nakayama, K., Imaki, H., Hirose, S., Jiang, Y., Abe, M., Tsukiyama, T., Nagahama, H., Ohno, S., Hatakeyama, S., Nakayama, K-I.: "Increased proliferation of B cells and autoimmune disease in protein kinase C-delta-deficient mice"Nature. (in press).

[文献書誌] Hiwasa, T., Nakata, M., Nakata, M., Ohno, S., Maki M., Suzuki, K., Takiguchi, M.: "Regulation of Transformed State by Calpastatin via PKCepsilon in NIH3T3 Mouse Fibroblasts"Biochem. Biophys. Res. Commun.. 290. 510-517 (2002)

[文献書誌] Harbers, M., Nomura, T., Ohno, S., Ishii, S.: "Intracellular localization of the ret finger protein depends on a functional nuclear export signal and protein kinase C activation"J. Biol Chem.. 276. 48596-48607 (2001)

[文献書誌] Okuda, H., Saitoh, K., Hirai, Si., S., Iwai, K., Takak Y., Baba, M., Minato, N., Ohno, S., Shuin, T.: "The Von Hippel-Lindau (VHL) tumor suppressor protein mediates ubiquitination of activated atypical protein kinase C"J Biol Chem.. 276. 43611-43617 (2001)

[文献書誌] Ohno, S.: "Intercellularjunctions and cellular polarity : the PAR-aPKC complex, a conserved core cassette playing fundamental roles in cell polarity"Curr. Opin. Cell Biol.. 13. 641-648 (2001)

[文献書誌] Zhao, X., Murata, T., Ohno, S., Day, N., Song, J., Nomura, N., Nakahara, T., Yokoyama, K.K.: "Protein kinase Ca plays a critical role in mannosylerythritol lipid-induced differentiation of melanoma B16 cells"J. Biol. Chem.. 276. 39903-39910 (2001)

[文献書誌] Ebnet, K., Suzuki, A., Horikoshi, Y., Hirose, T., Meyer, Zu., Brickwedde, M.K., Ohno, S., Vestweber, D.: "The cell polarity protein ASIP/PAR-3 directly associates with junctional adhesion molecule (JAM)"EMBO. J.. 20. 3738-3748 (2001)

[文献書誌] Yamaji, S., Suzuki, A., Sugiyama, Y., Koide, Y., Yoshida, M., Kanamori.H., Mohri, H., Ohno, S., Ishigatsubo, Y.: "Anovel integrin-linked kinase-binding protein, affixin, is involved in the early stage of cell-substrate interaction"J. Cell. Biol.. 153. 1251-1264 (2001)

[文献書誌] Takaki, Y., Hirai, S., Manabe, N., Izumi, Y., Hirose, T., Nakaya, M., Suzuki, A., Mizuno, K., Akimoto, K., Tsukita, S., Shuin, T., Ohno, S.: "Dynamic changes in protein components of the tight junction during liver regeneration"Cell Tissue Res.. 305. 399-409 (2001)

[文献書誌] Baba, M., Hirai, S., Kawakami, S., Kishida, T., Sakai, N., Kaneko, S., Yao, M., Shuin, T., Kubota, Y., Hosaka, M., Ohno, S.: "Tumor suppressor protein VHL is induced at high cell density and mediates contact inhibition of cell growth"Oncogene. 20. 2727-2736 (2001)

[文献書誌] Noda, Y., Takeya, R., Ohno, S., Naito, S., Ito, T., Sumimoto, H.: "Human homologues of the Caenorhabditis elegans cell polarity protein PAR6 as an adaptor that links the small GTPases Rac and Cdc42 to atypical protein kinase C"Genes Cells. 6. 107-119 (2001)

2001 年度実績報告書

大野茂男横浜市立大学, 医学部, 教授 (10142027)