ショウジョウバエの細胞分裂に伴うオルガネラ動態とメンブレントラフィックによる制御

2013 Fiscal Year Annual Research Report

• Project/Area Number: 23570004
• Research Institution: Kyoto Institute of Technology
• Principal Investigator: 井上 喜博 京都工芸繊維大学, 昆虫バイオメディカル教育研究センター, 准教授 (90201938)
• Co-Investigator(Kenkyū-buntansha): 山口 政光 京都工芸繊維大学, 工芸科学研究科, 教授 (00182460)
• Keywords: メンブレントラフィック / オルガネラ / 細胞質分裂 / ショウジョウバエ / 減数分裂

Research Abstract

ショウジョウバエ雄の減数分裂細胞は、サイズが大きく、分裂後期以降に微小管が果たす役割も解析できるという利点がある。各種細胞内構造の構成タンパクにGFP、RFP、YFP、Venus、CFPタグを付けたものを精母細胞で発現させ、それらの細胞内構造を同時に観察できるマルチカラーイメージング法を確立した。この方法により精母細胞をその成長期から減数第１、２分裂終了まで連続観察できる。蛍光タグを選べば複数の細胞内構造の変化を同時に観察できる。ミトコンドリア、ゴルジ体、小胞体の変化を明らかにした。この細胞にsiRNAを直接導入するelectroporation法も試みたが、効率が悪かった。代わりに、任意の遺伝子の２本鎖RNAとDcer2をGal4/UASシステムにより強制発現させることにより、同遺伝子を効率よくノックダウンできる系を構築した。系統センターからRNAi系統を入手すれば、ショウジョウバエのほぼ全遺伝子をノックダウンして、効果を調べることができる。本研究では、メンブレントラフィックに関わる遺伝子群1,030個の２本鎖RNAを発現させて、細胞質分裂に異常が表れる遺伝子を同定した（Kitazawa et al., 2014）。その結果、細胞内の小胞輸送を担うCOPIならびにCOPII小胞の構成因子が雄減数分裂細胞における細胞質分裂に必須なことがわかった。前者は細胞質分裂に必要な因子を分裂溝に運搬するために、後者はアクトミオシン環を細胞膜に固定するのに必要な因子を運搬するのに必要であった。一方、それらの小胞輸送因子が体細胞分裂にも関わるか検討した。幼虫期中枢神経系から神経芽細胞を調整して、その細胞分裂をタイムラプス観察した。同細胞特異的にCOPI, COPIIの構成因子の２本鎖RNAを発現させて効果を調べたが、雄減数分裂細胞でみられたような明確な細胞質分裂の欠損は観察されなかった。

Research Products

• [Journal Article] Orbit/CLASP is required for myosin accumulation at the cleavage furrow in Drosophila male meiosis. (2014)
  • Author(s): Kitazawa, D., Matsuo, T., Kaizuka, K., Miyauchi, C., Hayashi, D. and Inoue, Y. H.
  • Journal Title: PLoS ONE Volume: in press Pages: in press
  • Peer Reviewed
• [Journal Article] Drosophila MCM10 is required for DNA replication and differentiation in the compound eye. (2014)
  • Author(s): Vo, N., Taga, A., Inaba, Y., Hideki, Y., Cotterill, S. and Yamaguchi, M.
  • Journal Title: PLoS ONE Volume: 9 Pages: e93450.
  • DOI: 10.1371/journal.pone.0093450. eCollection 2014. PMID: 24686397
  • Peer Reviewed
• [Journal Article] DNA polymerase alpha interacts with PrSet7 and mediates H4K20 monomethylation in Drosophila. (2014)
  • Author(s): Sahashi, R., Crevel G., Pasko, J., Suyari, O., Nagai, R., Saura, M. M., Yamaguchi, M. and Cotterill, S.
  • Journal Title: J. Cell Science Volume: in press Pages: in press
  • Peer Reviewed
• [Journal Article] Identification of ter94, Drosophila VCP, as a strong modulator of motor neuron degeneration induced by knockdown of Caz, Drosophila FUS. (2014)
  • Author(s): Azuma, Y., Tokuda, T., Shimamura M., Kyotani, A., Sasayama H., Yoshida, T., Mizuta, I., Mizuno, T., Nakagawa, M., Fujikake, N., Ueyama, M., Nagai, Y. and Yamaguchi, M.
  • Journal Title: Hum. Mol. Genet. Volume: in press Pages: in press
  • Peer Reviewed
• [Journal Article] Drosophila myeloid leukemia factor acts with DREF to activate the JNK signaling pathway. (2014)
  • Author(s): anai, H., Yoshioka, Y., Yoshida, H., Nakao, Y., Plessis, A. and Yamaguchi, M.
  • Journal Title: Oncogenesis Volume: in press Pages: in press
  • Peer Reviewed
• [Journal Article] Metabolome analysis of Drosophila melanogaster during embryogenesis. (2014)
  • Author(s): Thuy An, P. N., Yamaguchi, M., Bamba, T. and Fukusaki, E.
  • Journal Title: PLos One Volume: in press Pages: in press
  • Peer Reviewed
• [Journal Article] dNF-YB plays dual roles in cell death and cell differentiation during Drosophila eye development. (2013)
  • Author(s): Ly, L.L., Suyari, O., Yoshioka, Y., Tue, N.T., Yoshida, H. and Yamaguchi, M.
  • Journal Title: Gene Volume: 520 Pages: 106-118
  • Peer Reviewed
• [Journal Article] Overexpression of Drosophila RFX3 induces apoptosis and interferes with differentiation of photoreceptor cells. (2013)
  • Author(s): Sugimoto, T., Ueda, T., Yoshida, H., Murakami, A. and Yamaguchi, M.
  • Journal Title: J. Carcinogene. Mutagene. Volume: 4 Pages: 2157-2518
  • Peer Reviewed
• [Journal Article] The Drosophila DOCK family protein Sponge is involved in differentiation of R7 photoreceptor cells. (2013)
  • Author(s): Eguchi, K., Yoshioka, Y., Yoshida, H., Morishita, K., Miyata, S., Hiai, H. and Yamaguchi, M.
  • Journal Title: Exp. Cell Res. Volume: 315 Pages: 2179-2195
  • Peer Reviewed
• [Journal Article] Temporal and spatial pattern of dref expression during Drosophila bristle development. (2013)
  • Author(s): Kawamori, A., Shimaji, K. and Yamaguchi, M.
  • Journal Title: Cell Struct. Funct. Volume: 38 Pages: 169-181
  • Peer Reviewed
• [Journal Article] Control of e2f1 and PCNA by Drosophila transcription factor DREF. (2013)
  • Author(s): Kawamori, A., Shimaji, K. and Yamaguchi, M.
  • Journal Title: Genesis Volume: 51 Pages: 741-750
  • Peer Reviewed
• [Journal Article] Genetic link between Heme Oxygenase and the signaling pathway of DNA damage in Drosophila melanogaster. (2013)
  • Author(s): Ida, H., Suyari, O., Shimamura, M., Tran, T. T., Yamaguchi, M. and Taketani, S.
  • Journal Title: Tohoku J. Exp. Med. Volume: 231 Pages: 117-125
  • Peer Reviewed
• [Journal Article] Functional analysis of Drosophila DNA polymerase ε p58 subunit. (2013)
  • Author(s): Sahashi, R., Matsuda, R., Suyari, O., Kawai, M., Yoshida, H., Cotterill, S. and Yamaguchi, M.
  • Journal Title: Am. J. Cancer Res. Volume: 3 Pages: 478-489
  • Peer Reviewed
• [Journal Article] Distribution and morphological changes of the Golgi apparatus during Drosophila spermatogenesis. (2013)
  • Author(s): Yasuno, Y., Inoue, Y. H. and Yamamoto, M.
  • Journal Title: Dev.Growth.Differ. Volume: 55 Pages: 635-647
  • Peer Reviewed
• [Presentation] Targeted induction of ER stress in Insulin-Producing Cells results in diabetes-like phenotype,inhibition of cell growth and dwarfism in Drosophila. (2013)
  • Author(s): Hinami, Y. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] 7SKsnRNP components such as LARP7 and 7SKsnRNP are dispensable for viability and fertility in Drosophila (2013)
  • Author(s): Okazaki, R. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] A role of mxc gene in larval lymph gland in Drosophila hematopoiesis (2013)
  • Author(s): Ozeki, T. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Sir2 can induce 4EBP and Catalase gene expression through Foxo activation in Drosophila. (2013)
  • Author(s): Nakahara, Y. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Identification of target genes of MXC protein, a tumor suppressor gene produ ct, and its transctiptional regulation in Drosophila. (2013)
  • Author(s): Awane, R. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Proper mitochondria dynamics of mitochondria is required for meiotic cell cycle progression in Drosophila spermatogenesis. (2013)
  • Author(s): Matsuo, T. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] COPII components are required for later stage of cytokinesis in Drosophila meiotic divisions (2013)
  • Author(s): Kaizuka, K. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Multiple role of Drosophila Mcm10 during compound eye development. (2013)
  • Author(s): Nicole Vo, Taga Ayano, Yasuhiro Inaba, Hideki Yoshida, Sue Cotterill, Masamitsu Yamaguchi
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Dynamics of epigenetic regulator jumonji (Jarid2) during Drosophila spermatogenesis. (2013)
  • Author(s): Narumi Toda, Nobuhiro Sasai, HIdeki Yoshida, Yoshihiro H. Inoue and Hiroshi Kimura, Masamitsu Yamaguchi
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] A Drosophila model for functional analysis of Organic Carrier Partner 1. (2013)
  • Author(s): Nguyen Tho Huu, Seiji Miyata, Hideki Yoshida, Masamitsu Yamaguchi
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Functional analysis of Drosophila DOCK-family Zizimin. (2013)
  • Author(s): Fumiko Ozawa, Kazushige Morishita, HIdeki Yoshida, Seiji Miyata, Masamitsu Yamaguchi
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Drosophila transcription factor DREF genetically interact with tumor suppressor genes. (2013)
  • Author(s): Yukari Terakata, Takeshi Horii, Shunsuke Fujiwara, Hideki Yoshida, Masamitsu Yamaguchi
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Cabeza that has been identified in Amyyotrophic Lateral Sclerosis regulates the EGFR signaling pathway in Drosophila. (2013)
  • Author(s): Akane Kyotani, Mai Shimomura, Yumiko Azuma, Takahiko Tokuda, Ikuko Mizuta, Masanari Nakagawa, Toshiki Mizuno, Hideki Yoshida, Masamitsu Yamaguchi
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
  • Year and Date: 20131203-20131206
• [Presentation] Nuclear envelope proteins, Lamins and nuclear pore complex are required for cytokinesis in Drosophila male meiosis. (2013)
  • Author(s): Hayashi, D. and Inoue, Y. H.
  • Organizer: EMBO Work Shop 2013 Drosophila Cell division cycle
  • Place of Presentation: Darlington, England
  • Year and Date: 20130912-20130916
  • Invited
• [Presentation] Orbit/CLASP is an essential factor for elongation that mature centrioles in Drosophila primary spermatocytes (2013)
  • Author(s): Asano, Y. and Inoue, Y. H.
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋、ウニンク愛知
  • Year and Date: 20130619-20130621
• [Presentation] 被覆小胞COPIIはショウジョウバエ雄減数分裂における細胞質分裂に必要である (2013)
  • Author(s): 貝塚加奈、井上喜博
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋、ウインク愛知
  • Year and Date: 20130619-20130621
• [Presentation] COPI-mediated membrane trafficking is required for cytokinesis in Drosophila male meiotic divisions (2013)
  • Author(s): Kitazawa, D. and Inoue, Y. H.
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋、ウインク愛知
  • Year and Date: 20130619-20130621
• [Presentation] ショウジョウバエ骨髄性白血病因子dMLFは転写因子DRFのコファクターとして働きJNK遺伝子を転写活性化する。 (2013)
  • Author(s): 梁井啓史、吉岡泰秀、吉田秀樹、山口政光
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋、ウインク愛知
  • Year and Date: 20130619-20130621
• [Presentation] 非古典的経路で小胞体へ標的化するmRNAの同定及び同機構解析系の構築 (2013)
  • Author(s): 梅河内隆成、越田大夢、山田百子、臼井一馬、佐藤哲也、宇山幹太、伊藤恵、大川恭介、山口政光、吉田秀樹
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋、ウインク愛知
  • Year and Date: 20130619-20130621
• [Presentation] DNA複製制御におけるショウジョウバエDNAポリメラーゼaとヒストンメチル化酵素Pr-Set7との相互作用 (2013)
  • Author(s): 佐橋律子、須鎗理、Crevel, G., Coterill, S. 山口政光
  • Organizer: 第６５回日本細胞生物学会大会
  • Place of Presentation: 名古屋、ウインク愛知
  • Year and Date: 20130619-20130621
• [Presentation] Developmental control of Drosophila spermatogenesis by Clasp/Orbit
  • Author(s): Inoue, Y. H.
  • Organizer: International symposium of IBRC in KIT
  • Place of Presentation: Kyoto
  • Invited
• [Presentation] Accumulation of ROS promotes muscle senescence in Drosophila adults and also influences muscle development.
  • Author(s): Hirai, J. and Inoue, Y. H.
  • Organizer: 第３６回日本分子生物学会年会
  • Place of Presentation: 神戸、国際会議場
• [Patent(Industrial Property Rights)] 抗老化物質のスクリーニング方法 (2014)
  • Inventor(s): 井上喜博
  • Industrial Property Rights Holder: 京都工芸繊維大学
  • Industrial Property Number: 特願2014-063045
  • Filing Date: 2014-03-26
  • Overseas