A genetic analysis on a novel regulatory mechanism to induce cell death in response to perturbation of dynamic instability of microtubules

• Project/Area Number: 17570006
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Genetics/Genome dynamics
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: INOUE Yoshihiro Kyoto Institute of Technology, Drosophila Genetic Resource Center, Lecturer, ショウジョウバエ遺伝資源センター, 講師 (90201938)
• Co-Investigator(Kenkyū-buntansha): YAMAGUCHI Masamitsu Kyoto Institute of Technology, Graduate School of Science and Technology, Professor, 工芸科学研究科, 教授 (00182460)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: microtubules / Drosophila / apoptosis / Orbit / dynamic instability

Research Abstract

Microtubules are indeed involved in many cellular phenomena. It has been shown that dynamic instability of microtubules is especially important for regulation of some cellular events. The Orbit/CLASP, a conserved microtubule-associated protein enriched at plus-ends of microtubules plays an essential role in regulation of the dynamic instability. Loss-of-function mutations of the orbit in Drosophila increase instability of microtubules. Conversely, over-expression of the gene in imaginal discs using GAL4-UAS system causes excessive stabilization of microtubules and consequently a formation of microtubule bundles that are never observed in normal cells at interphase. The over-expression of the orbit in eye disc cells after morphogenic furrows accumulates metaphase cells, suggesting it activates the microtubule assembly checkpoint in mitosis. We found that the over-expression eventually induces apoptosis dependent on Caspase 3 at post-mitotic stage in the eye discs. Knockdown of the gene also causes induction of the Caspase 3 in wing discs. This type of apoptosis cannot be observed in non-dividing cells such as larval salivary gland cells with over-expression of the orbit. The induction of apoptosis is required for a proper function of mitotic checkpoint genes and p53. Our data should suggest that cells with abnormal microtubules deviated from proper range of the dynamic instability could be recognized during mitosis and could be eliminated by the apoptosis from proliferating tissues.

Research Products

• [Journal Article] Transcriptional regulation of the Drosophila rfc1 gene by the DRE-DREF pathway. (2007)
  • Author(s): Tsuchiya, A., Inoue, Y.H., Ida, H., Kawase, Y., Okudaira, K., Ohno, K., Yoshida, H., Yamaguchi, M.
  • Journal Title: FEBS J. 274 Pages: 1818-1832
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Complex interference in the eye developmental pathway by Drosophila NF-YA. (2007)
  • Author(s): Yoshioka, Y., Suyari, O., Yamada M., Ohno, K., Hayashi, Y., Yamaguchi, M.
  • Journal Title: Genesis 45 Pages: 21-31
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system. (2007)
  • Author(s): Kim, Y.S, Shin, M.J., Yang, D.J., Yamaguchi, M., Park, S.Y., Yoo, M.A.
  • Journal Title: Genes to Cells. (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Prevalence of full-size P and KP elements in North America populations of Drosophila melanogaster. (2007)
  • Author(s): Itoh, M., Takeuchi, N., Yamaguchi, M., Yamamoto, M., Boussy, I., A.
  • Journal Title: Genetica (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of ZNF200 as a novel binding partner of histone H3 methyltransferase G9a. (2007)
  • Author(s): Nishida, M., Kato, M., Kato, Y., Sasai, N., Ueda, J., Tachibana, M., Shinkai, Y., Yamaguchi, M.
  • Journal Title: Genes to Cells. (in press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Transcriptional regulation of the Drosophila rfcl gene by the DRE-DREF pathway (2007)
  • Author(s): Tsuchiya, A., Inoue, Y.H., Ida, H., Kawase, Y., Okudaira, K., Ohno, K., Yoshida, H, Yamaguchi, M.
  • Journal Title: FEBS J. 274 Pages: 1818-1832
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Complex interference in the eye developmental pathway by Drosophila NF-YA. (2007)
  • Author(s): Yoshioka, Y, Suyari, O., Yamada, M., Ohno, K., Hayashi, Y., Yamaguchi, M.
  • Journal Title: Genesis. 45 Pages: 21-31
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Chromosome dynamics regulated by Orbit/CLASP protein family. (in Japanese) (2007)
  • Author(s): Ando, I., Suzuki.T, Inoue, Y.H.
  • Journal Title: NTS Press
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Transcriptional regulation of the Drosophila rfcl gene by the DRE-DREF pathway. (2007)
  • Author(s): Tsuchiya, A., Inoue, Y.H., Ida, H., Kawase, Y., Okudaira, K., Ohno, K., Yoshida, H., Yamaguchi, M.
  • Journal Title: FEBS J. 274 Pages: 1818-1832
• [Journal Article] Complex interference in the eye developmental pathway by Drosophila NF-YA. (2007)
  • Author(s): Yoshioka, Y., Suyari, O., Yamada, M., Ohno, K., Hayashi, Y., Yamaguchi, M.
  • Journal Title: Genesis 45 Pages: 21-31
• [Journal Article] Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system. (2007)
  • Author(s): Kim, Y.S, Shin, M.J., Yang, D.J., Yamaguchi, M., Park, S.Y., Yoo, M.A.
  • Journal Title: Genes to Cells (in press)
• [Journal Article] Identification of ZNF200 as a novel binding partner of histone H3 methyltransferase G9a. (2007)
  • Author(s): Nishida, M., Kato, M., Kato, Y., Sasai, N., Ueda, J., Tachibana, M., Shinkai, Y., Yamaguchi, M.
  • Journal Title: Genes to Cells (in press)
• [Journal Article] Drosophila Damaged DNA Binding Protein 1 contributes to genome stability in somatic cells. (2006)
  • Author(s): Shimanouchi, K., Takata, K., Yamaguchi, M., Murakami, S., Ishikawa, G., Takeuchi, R., Kanai, Y., Ruike, T., Nakamura, R., Abe, Y., Sakaguchi, K.
  • Journal Title: J. Biochem. 139 Pages: 51-58
  • NAID: 10018848289
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Antagonistic regulation of the Drosophila PCNA gene promoter by DREF and Cut. (2006)
  • Author(s): Seto, H., Hayashi, Y., Kwon, E., Taguchi, O., Yamaguchi, M.
  • Journal Title: Genes to Cells 11 Pages: 499-512
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genetic link between β-sarcoglycan and the Egfr signaling pathway. (2006)
  • Author(s): Hashimoto, R., Yamaguchi, M.
  • Journal Title: Biochem. Biophys. Res. Commun. 348 Pages: 212-221
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Drosophila Distal-less negatively regulates dDREF by inhibiting its DNA binding activity. (2006)
  • Author(s): Hayashi, Y., Kato, M., Seto, H., Yamaguchi, M.
  • Journal Title: Biochim. Biophys. Acta. 1759 Pages: 359-366
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of the Drosophila skpA gene as a novel target of the transcription factor DREF. (2006)
  • Author(s): Thao, D.T.P., Ida, H., Yoshida, H., Yamaguchi, M.
  • Journal Title: Exp. Cell Res. 312 Pages: 3641-3650
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Dynamic changes in the subcellular localization of Drosophila β-Sarcoglycan during the cell cycle. (2006)
  • Author(s): Hashimoto, R., Yamaguchi, M.
  • Journal Title: Cell Struct. Funct. 31 Pages: 173-180
  • NAID: 40015510810
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Drosophila Damaged DNA Binding Protein 1 contributes to genome stability in somatic cells. (2006)
  • Author(s): Shimanouchi, K., Takata, K., Yamaguchi, M., Murakami, S., Ishikawa, G, Takeuchi, R., Kanai, Y, Ruike, T, Nakamura, R Abe, Y.Sakaguchi, K.
  • Journal Title: J.Biochem. 139 Pages: 51-58
  • NAID: 10018848289
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic link between p-sarcoglycan and the Egfr signaling pathway. (2006)
  • Author(s): Hashimoto, R., Yamaguchi, M.
  • Journal Title: Biochem.Biophys.Res.Commun. 348 Pages: 212-221
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Drosophila Distal-less negatively regulates dDREF by inhibiting its DNA binding activity. (2006)
  • Author(s): Hayashi, Y., Kato, M., Seto, H.Yamaguchi, M.
  • Journal Title: Biochim.Biophys.Acta. 1759 Pages: 359-366
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dynamic changes in the subcellular localization of Drosophila β-Sarcoglycan during the cell cycle. (2006)
  • Author(s): Hashimoto, R., Yamaguchi, M.
  • Journal Title: Cell Struct.Funct. 31 Pages: 173-180
  • NAID: 40015510810
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic link between □-sarcoglycan and the Egfr signaling pathway. (2006)
  • Author(s): Hashimoto, R., Yamaguchi, M.
  • Journal Title: Biochem. Biophys. Res. Commun. 348 Pages: 212-221
• [Journal Article] Dynamic changes in the subcellular localization of Drosophila □-Sarcoglycan during the cell cycle. (2006)
  • Author(s): Hashimoto, R., Yamaguchi, M.
  • Journal Title: Cell Struct. Funct. 31 Pages: 173-180
• [Journal Article] Microtubule bundle formation and cell death induced by the human Orbit/CLASP N-terminal fragment. (2005)
  • Author(s): Aonuma, M., Miyamoto, M., Inoue, Y.H., et al.
  • Journal Title: Cell Struct.Funct. 20 Pages: 7-13
• [Journal Article] parva germina, a Gene Involved in germ cell maintenance during male and female Drosophila gametogenesis. (2005)
  • Author(s): Giovanna, M., Inoue, Y.H., Callaini, G
  • Journal Title: Dev.Dynamics 232 Pages: 835-844
• [Journal Article] Antagonistic regulation of the Drosophila PCNA gene promoter by DREF and Cut.
  • Author(s): Seto, H., Hayashi, Y, Kwon, E., Taguchi, O., Yamaguchi, M.
  • Journal Title: Genes to Cells 11 Pages: 499-512
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of the Drosophila skpA gene as a novel target of the transcription factor DREF.
  • Author(s): Thao, D.T.P., Ida, H., Yoshida, H.Yamaguchi, M.
  • Journal Title: Exp.Cell Res. 312 Pages: 3641-3650
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Transcriptional regulation of the Drosophila ANT gene by the DRE/DREF system.
  • Author(s): Kim, Y.S, Shin, M.J., Yang, D.J., Yamaguchi, M., Park, S.Y., Yoo, M.A.
  • Journal Title: Genes to Cells (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Prevalence of full-size P and KP elements in North America populations of Drosophila melanogaster.
  • Author(s): Itoh, M., Takeuchi, N., Yamaguchi, M., Yamamoto, M., Boussy, I., A.
  • Journal Title: Genetica (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of ZNF200 as a novel binding partner of histone H3 methyltransferase G9a.
  • Author(s): Nishida, M., Kato, M., Kato, Y., Sasai, N., Ueda, J., Tachibana, M., Shinkai, Y., Yamaguchi, M.
  • Journal Title: Genes to Cells (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] -日本の遣伝学の潮流-Orbit/Claspタンパクファミリーによるショウジョウバエの染色体ダナミックス (2007)
  • Author(s): 安藤至, 鈴木隆夫, 井上喜博
  • Total Pages: 340
  • Publisher: NTS出版
  • Description: 「研究成果報告書概要(和文)」より
• [Book] -日本の遺伝学の潮流-Orbit/Claspタンパクファミリーによるショウジョウバエの染色体ダイナミックス (2007)
  • Author(s): 安藤至, 鈴木隆夫, 井上喜博
  • Total Pages: 340
  • Publisher: NTS出版