Coupling of membrane traffic and cytoskeleton remodeling in the regulation of cell motility

2012 Fiscal Year Final Research Report

• Project/Area Number: 22390013
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Biological pharmacy
• Research Institution: Kyoto University
• Principal Investigator: NAKAYAMA Kazuhisa 京都大学, 大学院・薬学研究科, 教授 (40192679)
• Project Period (FY): 2010 – 2012
• Keywords: メンブレントラフィック / 小胞輸送 / 細胞骨格 / 細胞運動 / 細胞分裂

Research Abstract

Cell motility and cell division are under coordinated regulation of membrane traffic and actin cytoskeleton remodeling. The Arf and Rab families of small GTPases regulate membrane traffic, and the Rho family small GTPases regulate cytoskeleton remodeling. In this study, we showed that Arfaptins, which are dual effectors of Arl1 and Rac1, are recruited onto trans-Golgi membranes through interacting with Arl1, and induce membrane tubulation in an Arl1-dependent manner. X-ray crystal structure of the Arl1-Arfaptin-2 complex revealed the molecular basis for the membrane recruitment of Arfaptin-2 and indicated a possibility that Arl1 and Rac1 compete with each other for binding to Arfaptin-2.On the other hand, we showed that Arf6 is recruited to the Flemming body during cytokinesis through interacting with MLKP1, which forms a complex with MgcRacGAP, and is involved in regulation of cytokinesis. Furthermore, We revealed the structure of the Arf6-MKLP1 complex by X-ray crystallography and suggested the molecular basis for recruitment of Arf6 onto the Flemming body. We further showed that EFA6, an Arf-GEF, is involved in local activation of Arf6 during cytokinesis.

Research Products

• [Journal Article] EFA6 activates Arf6 and participates in its targeting to the Flemming body during cytokinesis. (2013)
  • Author(s): Ueda, T., Hanai, A., Takei, T., Kubo, K.,Ohgi, M., Sakagami, H., Takahashi, S.,Shin, H.-W. & Nakayama, K
  • Journal Title: FEBS Lett Volume: 587 Pages: 1617-1623.
  • DOI: doi:10.1016/j.febslet.2013.03.042.
  • Peer Reviewed
• [Journal Article] Mitosis-coupled, microtubule-dependent clustering of endosomal vesicles around centrosomes. (2013)
  • Author(s): Takatsu, H., Katoh, Y., Ueda, T., Waguri, S., Murayama, T., Takahashi, S., Shin, H.-W. &Nakayama, K.
  • Journal Title: Cell Struct. Funct Volume: 38 Pages: 31-41
  • DOI: doi:10.1247/csf.12028.
  • Peer Reviewed
• [Journal Article] ARF1 and ARF3 are required for the integrity of recycling endosomes and the recycling pathway (2012)
  • Author(s): Kondo, Y., Hanai, A., Nakai, W., Katoh, Y.,Nakayama, K. & Shin, H.-W.
  • Journal Title: ARF1 and ARF3 are required for the integrity of recycling endosomes and the recycling pathway Volume: 37 Pages: 141-154
  • DOI: doi:10.1247/csf.12015.
  • Peer Reviewed
• [Journal Article] Structural basis for membrane binding specificity of the Bin/Amphiphysin/Rvs (BAR) domain of Arfaptin-2 determined by Arl1 GTPase. (2012)
  • Author(s): Nakamura, K., Man, Z., Xie, Y., Hanai, A., Makyio, H., Kawasaki, M., Kato, R., Shin, H.-W., Nakayama, K. & Wakatsuki, S.
  • Journal Title: Biol. Chem. Volume: 287 Pages: 25478-25489.
  • DOI: doi:10.1074/jbc.M112.365783.
  • Peer Reviewed
• [Journal Article] Rab11 regulates exocytosis of recycling vesicles at the plasma membrane. (2012)
  • Author(s): Takahashi, S., Kubo, K., Waguri, S., Yabashi, A., Shin, H.-W., Katoh, Y. & Nakayama, K.
  • Journal Title: J. Cell Sci., 1 Volume: 125 Pages: 4049-4057
  • DOI: doi: 10.1242/jcs.102913.
  • Peer Reviewed
• [Journal Article] Structural basis for Arf6-MKLP1complex formation on the Flemming body responsible for cytokinesis. (2012)
  • Author(s): Makyio, H., Ohgi, M., Takei, T., Takahashi, S., Takatsu, H., Katoh, Y., Hanai, A., Ueda, T., Kanaho, Y., Xie, Y., Shin, H.-W., Kamikubo, H., Kataoka, M., Kawasaki, M., Kato, R., Wakatsuki, S. & Nakayama, K.
  • Journal Title: EMBO J Volume: 31 Pages: 2590-2603
  • DOI: doi:10.1038/emboj.2012.89
  • Peer Reviewed
• [Journal Article] Shigella targets epithelial tricellular junctions and uses a noncanonical clathrin-dependent endocytic pathway to spread between cells. (2012)
  • Author(s): Fukumatsu, M., Ogawa, M., Arakawa, S.,Suzuki, M., Furuse, M., Nakayama, K., Shimizu, S., Kim, M., Mimuro, H. & Sasakawa, C.
  • Journal Title: Cell Host Microbe Volume: 11 Pages: 325-336
  • DOI: doi:10.1016/j.chom.2012.03.001.
  • Peer Reviewed
• [Journal Article] Restriction of mast cell proliferation through hyaluronan synthesis by co-cultured fibroblasts. (2012)
  • Author(s): Takano, H., Furuta, K., Yamashita, K., Sakanaka, M., Itano, N., Gohda, E., Nakayama, K., Kimata, K., Sugimoto, Y., Ichikawa, A. & Tanaka, S.
  • Journal Title: Biol. Pharm. Bull. Volume: 35 Pages: 408-412
  • DOI: doi:10.1248/bpb.35.408.
  • Peer Reviewed
• [Journal Article] GBF1-Arf-COPI-ArfGAP-mediated Golgi-to-ER transport involved in regulation of lipid homeostasis. (2011)
  • Author(s): Takashima, K., Saitoh, A., Hirose, S., Nakai, W., Kondo, Y., Takasu, Y., Kakeya, H., Shin, H.-W. & Nakayama, K8.
  • Journal Title: Cell Struct. Funct Volume: 36 Pages: 223-235
  • DOI: doi:10.1247/csf.11035.
  • Peer Reviewed
• [Journal Article] ATP9B, a P4-ATPase (a putative aminophospholipid translocase), localizes to the trans-Golgi network in a CDC50-independent manner. (2011)
  • Author(s): Takatsu, H., Baba, K., Shima, T., Umino, H., Kato, U., Umeda, M., Nakayama, K. & Shin, H.-W.
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 38159-38167.
  • DOI: doi:10.1074/jbc.M111.281006.
  • Peer Reviewed
• [Journal Article] Distinct roles of Rab11 and Arf6 in the regulation of Rab11-FIP3/Arfophilin-1 localization in mitotic cells (2011)
  • Author(s): Takahashi, S., Takei, T., Koga, H., Takatsu,H., Shin, H.-W. & Nakayama, K.
  • Journal Title: Genes Cells Volume: 16 Pages: 938-950
  • DOI: doi:10.1111/j.1365-2443.2011.01538.x.
  • Peer Reviewed
• [Journal Article] Arfaptins are localized to the trans-Golgi by interaction with Arl1, but not Arfs. (2011)
  • Author(s): Man, Z., Kondo, Y., Koga, H., Umino, H., Nakayama, K. & Shin, H.-W.
  • Journal Title: J. Biol. Chem. Volume: 286 Pages: 11569-11578
  • DOI: doi:10.1074/jbc.M110.201442.
  • Peer Reviewed
• [Presentation] 細胞分裂におけるメンブレントラフィックの役割:細胞生物学と構造生物学の融合研究 (2012)
  • Author(s): 中山和久
  • Organizer: 第1回物構研サイエンスフェスタ
  • Place of Presentation: つくば
  • Year and Date: 2012-03-15
• [Presentation] 細胞分裂時のオルガネラの分配の調節. (2011)
  • Author(s): 中山和久
  • Organizer: 千里ライフサイエンスセミナー「生命科学を支えるオルガネラ研究の新展開」
  • Place of Presentation: 豊中
  • Year and Date: 2011-09-30
• [Presentation] ゴルジ体に局在する低分子量 GTPase ARF の機能の再考 (2011)
  • Author(s): 申 惠媛、中井和香、近藤由美香、花井綾子、中山和久
  • Organizer: 第84回日本生化学会大会シンポジウム
  • Place of Presentation: 京都
  • Year and Date: 2011-09-23
• [Presentation] Mitosis-Coupled Sequestration of Recycling Endosomes around Spindle Pole Centrosomes (2010)
  • Author(s): Nakayama, K., Takatsu, H., Ueda, T., Waguri, S., Takahashi, S., Kondo, Y. & Shin, H.-W.
  • Organizer: 第33回日本分子生物学会年会・第83回日本生化学会大会合同大会シンポジウム
  • Place of Presentation: 神戸
  • Year and Date: 2010-12-09
• [Presentation] Regulation of Arf6 and Rab11 during cell division. FASEB Summer Research Conference (2010)
  • Author(s): Nakayama, K
  • Organizer: Arf Family G Proteins
  • Place of Presentation: Carefree, Arizona, USA.
  • Year and Date: 2010-08-18
• [Remarks]
  • URL: http://www.pharm.kyoto-u.ac.jp/physchem/