Checkpoint analyses using anti -phosphoepitope-specific antibody

2011 Fiscal Year Final Research Report

• Project Area: Cell Proliferation Control
• Project/Area Number: 19057010
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Aichi Cancer Center Research Institute
• Principal Investigator: INAGAKI Masaki 愛知県がんセンター(研究所), 発がん制御研究部, 部長 (30183007)
• Co-Investigator(Kenkyū-buntansha): NAKANISHI Makoto 名古屋市立大学, 医学(系)研究科(研究院), 教授 (40217774)
• Project Period (FY): 2007 – 2011
• Keywords: Chk1 / チェックポイント / リン酸化修飾 / 中心体 / G0/G1移行 / トリコプレイン

Research Abstract

Chk1 is a protein kinase implicated in DNA maintenance checkpoint. Chk1 function is regulated through the phosphorylation of its regulatory domain. However, only ATR was long believed to functionally regulate Chk1. In this study, we have demonstrated that a variety of protein kinases phosphorylate and thereby regulate Chk1. In addition, we have also shown novel Chk1 function at the G0/G1 transition.

Research Products

• [Journal Article] Trichoplein and Aurora A block aberrant primary cilia assembly in proliferating cells (2012)
  • Author(s): Inoko, A., Matsuyama, M., Goto, H., Ohmuro-Matsuyama, Y., Hayashi, Y., Enomoto, M., Ibi, M., Urano, T., Yonemura, S., Kiyono, T., Izawa, I. Inagaki, M.
  • Journal Title: J. Cell Biol. Volume: 197 Pages: 391-405
  • DOI: doi:10.1083/jcb.201106101
  • Peer Reviewed
• [Journal Article] P90 RSK arranges Chk1 in the nucleus for monitoring of genomic integrity during cell proliferation (2012)
  • Author(s): Li, P., Goto, H., Kasahara, K., Matsuyama, M., Wang, Z., Yatabe, Y., Kiyono, T., Inagaki, M.
  • Journal Title: Mol. Biol. Cell Volume: 23 Pages: 1582-1592
  • DOI: doi:10.1091/mbc.E11-10-0883
  • Peer Reviewed
• [Journal Article] Trichoplein controls microtubule anchoring at the centrosome by binding to Odf2 and ninein (2011)
  • Author(s): Ibi, M., Zou, P., Inoko, A., Shiromizu, T., Matsuyama, M., Hayashi, Y., Enomoto, M., Mori, D., Hirotsune, S., Kiyono, T., Tsukita, S., Goto, H.. Inagaki, M.
  • Journal Title: J. Cell Sci. Volume: 124 Pages: 857-864
  • DOI: doi:10.1242/jcs.075705
  • Peer Reviewed
• [Journal Article] Vimentin organization modulates the formation of lamellipodia (2011)
  • Author(s): Helfand, B. T., Mendez, M. G., Murthy, S. N., Shumaker, D. K., Grin, B., Mahammad, S., Aebi, U., Wedig, T., Wu, Y. I., Hahn, K. M., Inagaki, M., Herrmann, H., Goldman, R. D.
  • Journal Title: Mol. Biol. Cell Volume: 22 Pages: 1274-1289
  • DOI: doi:10.1091/mbc.E10-08-0699
  • Peer Reviewed
• [Journal Article] Nuclear Chk1 prevents premature mitotic entry (2011)
  • Author(s): Matsuyama, M., Goto, H., Kasahara, K., Kawakami, Y., Nakanishi, M., Kiyono, T., Goshima, N., Inagaki, M.
  • Journal Title: J. Cell Sci. Volume: 124 Pages: 2113-2119
  • DOI: doi:10.1242/ jcs.086488
  • Peer Reviewed
• [Journal Article] DNA lesions induced by replication stress trigger mitotic aberration and tetraploidy development (2010)
  • Author(s): Ichijima, Y., Yoshioka, K., Yoshioka, Y., Shinohe, K., Fujimori, H., Unno, J., Takagi, M., Goto, H., Inagaki, M., Mizutani, S., Teraoka, H.
  • Journal Title: PLoS ONE Volume: 5 Pages: e8821
  • DOI: doi:10.1371/journal.pone.0008821
  • Peer Reviewed
• [Journal Article] Withaferin A targets intermediate filaments glial fibrillary acidic protein and vimentin in a model of retinal gliosis (2010)
  • Author(s): Bargagna-Mohan, P., Paranthan, R. R., Hamza, A., Dimova, N., Trucchi, B., Srinivasan, C., Elliott, G. I., Zhan, C. G., Lau, D. L., Zhu, H., Kasahara, K., Inagaki, M., Cambi, F., Mohan, R.
  • Journal Title: J. Biol. Chem. Volume: 285 Pages: 7657-7669
  • DOI: doi:10.1074/jbc.M109.093765
  • Peer Reviewed
• [Journal Article] Association between vitamin D and calcium intake and breast cancer risk according to menopausal status and receptor status in Japan (2010)
  • Author(s): Kawase, T., Matsuo, K., Suzuki, T., Hirose, K., Hosono, S., Watanabe, M., Inagaki, M., Iwata, H., Tanaka, H., Tajima, K.
  • Journal Title: Cancer Sci. Volume: 101 Pages: 1234-1240
  • DOI: doi:10.1111/j.1349-7006.2010.01496.x
  • Peer Reviewed
• [Journal Article] 14-3-3γ mediates Cdc25A proteolysis to block premature mitotic entry after DNA damage (2010)
  • Author(s): Kasahara, K., Goto, H., Enomoto, M., Tomono, Y., Kiyono, T., Inagaki, M.
  • Journal Title: EMBO J. Volume: 29 Pages: 2802-2812
  • DOI: doi:10.1038/emboj.2010.157
  • Peer Reviewed
• [Journal Article] Novel positive feedback loop between Cdk1 and Chk1 in the nucleus during the G2/M transition (2009)
  • Author(s): Enomoto, M., Goto, H., Tomono, Y., Kasahara, K., Tsujimura, K., Kiyono, T., Inagaki, M.
  • Journal Title: J. Biol. Chem. Volume: 284 Pages: 34223-34230
  • DOI: doi:10.1074/jbc.C109.051540
  • Peer Reviewed
• [Journal Article] Non-pathogenic protein aggregates in skeletal muscle in MLF1 transgenic mice (2008)
  • Author(s): Li, Z. F., Wu, X., Jiang, Y., Liu, J., Wu, C., Inagaki, M., Izawa, I., Mizisin, A. P., Engvall, E., Shelton, G. D.
  • Journal Title: J. Neurol. Sci. Volume: 264 Pages: 77-86
  • DOI: 10.1016/j.jns.2007.07.027
  • Peer Reviewed
• [Journal Article] Protein phosphatase 4 catalytic subunit regulates Cdk1 activity and microtubule organization via NDEL1 dephosphorylation (2008)
  • Author(s): Toyo-oka, K., Mori, D., Yano, Y., Shiota, M., Iwao, H., Goto, H., Inagaki, M., Hiraiwa, N., Muramatsu, M., Wynshaw-Boris, A., Yoshiki, A., Hirotsune, S.
  • Journal Title: J. Cell Biol. Volume: 180 Pages: 1133-1147
  • DOI: doi: 10.1083/jcb.200705148
  • Peer Reviewed
• [Journal Article] Palmitoylation of ERBIN is required for its plasma membrane localization (2008)
  • Author(s): Izawa, I., Nishizawa, M., Hayashi, Y., Inagaki, M.
  • Journal Title: Genes Cells Volume: 13 Pages: 691-701
  • DOI: doi:10.1111/j.1365-2443.2008.01198.x
  • Peer Reviewed
• [Journal Article] eIF3k regulates apoptosis in epithelial cells by releasing caspase 3 from keratin-containing inclusions (2008)
  • Author(s): Lin, Y. M., Chen, Y. R., Lin, J. R., Wang, W. J., Inoko, A., Inagaki, M., Wu, Y. C., Chen, R. H.
  • Journal Title: J. Cell Sci. Volume: 121 Pages: 2382-2393
  • DOI: doi:10.1242/ jcs.021394
  • Peer Reviewed
• [Journal Article] The keratin-binding protein Albatross regulates polarization of epithelial cells (2008)
  • Author(s): Sugimoto, M., Inoko, A., Shiromizu, T., Nakayama, M., Zou, P., Yonemura, S., Hayashi, Y., Izawa, I., Sasoh, M., Uji, Y., Kaibuchi, K., Kiyono, T., Inagaki, M.
  • Journal Title: J. Cell Biol. Volume: 183 Pages: 19-28
  • DOI: doi:10.1083/jcb.200803133
  • Peer Reviewed
• [Journal Article] Chk1 phosphorylation at Ser286 and Ser301 occurs with both stalled DNA replication and damage checkpoint stimulation (2008)
  • Author(s): Ikegami, Y., Goto, H., Kiyono, T., Enomoto, M., Kasahara, K., Tomono, Y., Tozawa, K., Morita, A., Kohri, K., Inagaki, M.
  • Journal Title: Biochem. Biophys. Res. Commun. Volume: 337 Pages: 1227-1231
  • DOI: 10.1016/j.bbrc.2008.10.119
  • Peer Reviewed
• [Journal Article] Production of a site- and phosphorylation state-specific antibody (2007)
  • Author(s): Goto, H. and Inagaki, M.
  • Journal Title: Nat. Protoc. Volume: 2 Pages: 2574-2581
  • DOI: doi:10.1038/nprot.2007.374
  • Peer Reviewed
• [Journal Article] Role of phosphorylation on the structural dynamics and function of types III and IV intermediate filaments (2007)
  • Author(s): Sihag, R. K., Inagaki, M., Yamaguchi, T., Shea, T. B., Pant, H. C.
  • Journal Title: Exp. Cell Res. Volume: 313 Pages: 2098-2109
  • DOI: 10.1016/j.yexcr.2007.04.010
  • Peer Reviewed
• [Journal Article] Inhibition of Aurora-B function increases formation of multinucleated cells in p53 gene deficient cells and enhances anti-tumor effect of temozolomide in human glioma cells (2007)
  • Author(s): Tsuno, T., Natsume, A., Katsumata, S., Mizuno, M., Fujita, M., Osawa, H., Nakahara, N., Wakabayashi, T., Satoh, Y., Inagaki, M., Yoshida, J.
  • Journal Title: J. Neurooncol. Volume: 83 Pages: 249-258
  • DOI: doi:10.1007/s11060-007-9335-1
  • Peer Reviewed
• [Journal Article] Novel regulation of checkpoint kinase 1 : Is checkpoint kinase 1 a good candidate for anti-cancer therapy?
  • Author(s): Goto, H., Izawa, I., Li, P. and Inagaki, M.
  • Journal Title: Cancer Sci. Volume: (in press)
  • DOI: doi:10.1111/j.1349-7006.2012.02280.x
  • Peer Reviewed
• [Presentation] Novel regulatory mechanism of Plk1 at mitosis (2011)
  • Author(s): Inagaki, M.
  • Organizer: MEXT Priority Research Project International Symposium "Cell Division"
  • Place of Presentation: Hakone
  • Year and Date: 2011-06-29
• [Presentation] Anti-phospho peptide antibodies-a tool to address the regulatory function of intermediate filaments and cell cycle progression in astrocytes (2009)
  • Author(s): Inagaki, M.
  • Organizer: the 6th European Conference on Intermediate Filaments (Nanofilaments) in Health and Disease
  • Place of Presentation: Sweden
  • Year and Date: 2009-06-20
• [Presentation] Signaling and cell architecture (2009)
  • Author(s): Inagaki, M.
  • Organizer: Receptor Program Ad Hoc Seminar
  • Place of Presentation: Turku
  • Year and Date: 2009-06-15
• [Presentation] Cell cycle and architecture (2009)
  • Author(s): Inagaki, M.
  • Organizer: Regulation and Manipulation of Information Flow within Dynamic Protein and Lipid Environments (SFB 645 Workshop Cell Architecture and Cell Signalling)
  • Place of Presentation: Bonn
  • Year and Date: 2009-06-12
• [Presentation] Trichoplein regulates microtubule anchoring and suppresses a cilia assembly program at the mother centriole (2009)
  • Author(s): Inagaki, M.
  • Organizer: Global Center of Excellence (COE) Program 1st International Symposium "Signaling of Cancer Cells"
  • Place of Presentation: Nagoya
  • Year and Date: 2009-01-23
• [Book] Detection of Protein Phosphorylation by Open-Sandwich Immunoassay. Integrative Proteomics. ed. Leung, H. -C. E. (2012)
  • Author(s): Ohmuro-Matsuyama, Y., Inagaki M., Ueda, H.
  • Total Pages: 197-214
  • Publisher: InTech (ISBN 978-953-51-0070-6)
• [Book] 「チェックポイントキナーゼ1 (Chk1)の新規制御機構とがん」 シグナル伝達研究最前線2012実験医学増刊 Vol.30-N0.5 井上純一郎, 武川睦寛, 徳川文稔, 今井浩三(編集) (2012)
  • Author(s): 後藤英仁, 稲垣昌樹
  • Total Pages: 106-110
  • Publisher: 羊土社
• [Book] 細胞周期フロンティア (2010)
  • Author(s): 佐方功幸, 稲垣昌樹, 岸本健雄(編)
  • Total Pages: 268
  • Publisher: 共立出版
• [Book] 「G2/M移行期におけるCDK1の活性化およびチェックポイント解除機構」細胞周期フロンティア (2010)
  • Author(s): 後藤英仁, 稲垣昌樹
  • Total Pages: 52-57
  • Publisher: 共立出版
• [Book] 「抗リン酸化ペプチド抗体と細胞周期研究」細胞周期フロンティア (2010)
  • Author(s): 笠原広介, 稲垣昌樹
  • Total Pages: 5-10
  • Publisher: 共立出版
• [Book] 「細胞周期研究の現状と展望」細胞周期フロンティア (2010)
  • Author(s): 稲垣昌樹, 佐方功幸
  • Total Pages: 248-250
  • Publisher: 共立出版
• [Book] リン酸化抗体と細胞周期 細胞工学Vol.28 No.1 (2009)
  • Author(s): 大室(松山)有紀, 稲垣昌樹
  • Total Pages: 50-51
  • Publisher: 秀潤社
• [Book] Close UP実験法 特定部位の翻訳後修飾を特異的に認識する.抗体作製法-抗リン酸化(ペプチド)抗体作製法, 実験医学, 26巻, 18号 (2008)
  • Author(s): 後藤英仁, 稲垣昌樹
  • Total Pages: 2965-2972
  • Publisher: 羊土社
• [Book] リン酸化-特集蛋白質修飾- 分子細胞治療, 6巻, 1号 (2007)
  • Author(s): 後藤英仁, 稲垣昌樹
  • Total Pages: 4-9
  • Publisher: 先端医学社
• [Book] 「中間径フィラメント」生化学事典
  • Author(s): 衣斐美歩, 稲垣昌樹
  • Publisher: 朝倉書店(in press)
• [Remarks]
  • URL: http://www.pref.aichi.jp/cancer-center/ri/01bumon/08hatsugan_seigyo/index.html