Elucidation of the molecular mechanism of Rab35-activation-dependent neurite outgrowth

• Project/Area Number: 15H04367
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Cell biology
• Research Institution: Tohoku University
• Principal Investigator: Fukuda Mitsunori 東北大学, 生命科学研究科, 教授 (50311361)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000); Fiscal Year 2017: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2016: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2015: ¥6,500,000 (Direct Cost: ¥5,000,000、Indirect Cost: ¥1,500,000)
• Keywords: 低分子量G蛋白質Rab / 神経突起伸長 / 小胞輸送 / 神経成長因子 / スクリーニング / Rabカスケード / Rabエフェクター / グアニンヌクレオチド交換因子 / 膜輸送

Outline of Final Research Achievements

Neurite outgrowth, which is a prerequisite for neuronal network formation, requires massive addition of proteins and lipids to the tips of growing neurites by membrane trafficking. Accumulating evidence has suggested that several Rab small GTPases such as Rab8 and Rab35 on recycling endosomes promote neurite outgrowth, but their regulatory mechanisms during neurite outgrowth are poorly understood. In this study, we identified DENND1A as a Rab35-GEF and Rabin8 as a Rab8-GEF and found that both GEFs are required for neurite outgrowth of PC12 cells. Knockdown of Rabin8 resulted in inhibition of neurite outgrowth, whereas its overexpression promoted it. We also found that Rabin8 is recruited to recycling endosomes in a Rab11-dependent manner and that it activates Rab8 and Rab10 there during neurite outgrowth. Furthermore, we comprehensively screened TBC proteins/Rab-GAPs and Rabs and succeeded in identifying TBC1D12 and Rab20 as novel negative regulators of neurite outgrowth.

Research Products

• [Int'l Joint Research] Tel Aviv University(Israel)
• [Int'l Joint Research] University of Oxford(英国)
• [Int'l Joint Research] Universidad de Chile(Chile)
• [Int'l Joint Research] University of Oxford(英国)
• [Int'l Joint Research] Tel Aviv University(Israel)
• [Journal Article] Rab20, a novel Rab small GTPase that negatively regulates neurite outgrowth of PC12 cells. (2018)
  • Author(s): Oguchi, M. E., Etoh, K. and Fukuda, M.
  • Journal Title: Neurosci. Lett. Volume: 662 Pages: 324-330
  • DOI: 10.1016/j.neulet.2017.10.056
  • Peer Reviewed / Open Access
• [Journal Article] TBC1D12 is a novel Rab11-binding protein that modulates neurite outgrowth of PC12 cells. (2017)
  • Author(s): Oguchi, M. E., Noguchi, K. and Fukuda, M.
  • Journal Title: PLoS One Volume: 12 Issue: 4 Pages: e0174883-e0174883
  • DOI: 10.1371/journal.pone.0174883
  • Peer Reviewed / Open Access
• [Journal Article] The GTPase Rab43 controls the anterograde ER-Golgi trafficking and sorting of GPCRs. (2017)
  • Author(s): Li, C., Wei, Z., Fan, Y., Huang, W., Su, Y., Li, H., Dong, Z., Fukuda, M., Khater, M. and Wu, G.
  • Journal Title: Cell Rep. Volume: 21 Issue: 4 Pages: 1089-1101
  • DOI: 10.1016/j.celrep.2017.10.011
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rab5 is critical for SNAP23 regulated granule-granule fusion during compound exocytosis. (2017)
  • Author(s): Klein, O., Roded, A., Zur, N., Azouz, N. P., Pasternak, O., Hirschberg, K., Hammel, I., Roche, P. A., Yatsu, A., Fukuda, M., Galli, S. J. and Sagi-Eisenberg, R.
  • Journal Title: Sci. Rep. Volume: 7 Issue: 1 Pages: 15315-15315
  • DOI: 10.1038/s41598-017-15047-8
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Rab32 subfamily small GTPases: pleiotropic Rabs in endosomal trafficking. (2017)
  • Author(s): Ohbayashi, N., Fukuda, M. and Kanaho, Y.
  • Journal Title: J. Biochem. Volume: 62 Issue: 2 Pages: 65-71
  • DOI: 10.1093/jb/mvx027
  • Peer Reviewed
• [Journal Article] Cdk5 regulation of the GRAB-mediated Rab8-Rab11 cascade in axon outgrowth. (2017)
  • Author(s): Furusawa, K., Asada, A., Urrutia, P., Gonzalez-Billault, C., Fukuda, M. and Hisanaga, S. I.
  • Journal Title: J. Neurosci. Volume: 37 Issue: 4 Pages: 790-806
  • DOI: 10.1523/jneurosci.2197-16.2017
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] M-INK, a novel tool for visualizing melanosomes and melanocores. (2017)
  • Author(s): Ishida, M., Marubashi, S. and Fukuda, M.
  • Journal Title: J. Biochem. Volume: 161 Pages: 323-326
  • DOI: 10.1093/jb/mvw100
  • NAID: 40021219775
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] C9orf72 and RAB7L1 regulate vesicle trafficking in amyotrophic lateral sclerosis and frontotemporal dementia. (2017)
  • Author(s): Aoki, Y., Manzano, R., Lee, Y., Dafinca, R., Aoki, M., Douglas, A. G. L., Varela, M. A., Sathyaprakash, C., Scaber, J., Barbagallo, P., Vader, P., Mager, I., Ezzat, K., Turner, M. R., Ito, N., Gasco, S., Ohbayashi, N., El-Andaloussi, S., Takeda, S., Fukuda, M., Talbot, K. and Wood, M. J. A.
  • Journal Title: Brain Volume: 140 Issue: 4 Pages: 887-897
  • DOI: 10.1093/brain/awx024
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Parkin promotes proteasomal degradation of synaptotagmin IV by accelerating polyubiquitination. (2017)
  • Author(s): Kabayama, H., Tokushige, N., Takeuchi, M., Kabayama, M., Fukuda M. and Mikoshiba, K.
  • Journal Title: Mol. Cell. Neurosci. Volume: 80 Pages: 89-99
  • DOI: 10.1016/j.mcn.2017.02.006
  • Peer Reviewed
• [Journal Article] P53- and mevalonate pathway-driven malignancies require Arf6 for metastasis and drug resistance. (2016)
  • Author(s): Hashimoto, A., Oikawa, T., Hashimoto, S., Sugino, H., Yoshikawa, A., Otsuka, Y., Handa, H., Onodera, Y., Nam, J.M., Oneyama, C., Okada, M., Fukuda M. and Sabe, H.
  • Journal Title: J Cell Biol. Volume: 213 Issue: 1 Pages: 81-95
  • DOI: 10.1083/jcb.201510002
  • NAID: 120005763128
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Regulation of podocalyxin trafficking by Rab small GTPases in 2D and 3D epithelial cell cultures. (2016)
  • Author(s): Mrozowska, P. S. and Fukuda, M.
  • Journal Title: J. Cell Biol. Volume: 213 Issue: 3 Pages: 355-369
  • DOI: 10.1083/jcb.201512024
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] A Varp-binding protein, RACK1, regulates dendrite outgrowth through stabilization of Varp protein in mouse melanocytes. (2016)
  • Author(s): Marubashi, S., Ohbayashi, N. and Fukuda, M.
  • Journal Title: J. Invest. Dermatol. Volume: 136 Issue: 8 Pages: 1672-1680
  • DOI: 10.1016/j.jid.2016.03.034
  • NAID: 120007129451
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Rabin8 regulates neurite outgrowth in both GEF-activity-dependent and -independent manners. (2016)
  • Author(s): Homma, Y. and Fukuda, M.
  • Journal Title: Mol. Biol. Cell Volume: 27 Issue: 13 Pages: 2107-2118
  • DOI: 10.1091/mbc.e16-02-0091
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Rab35 functions in axon elongation are regulated by p53-related protein kinase (PRPK) in a mechanism that involves Rab35 protein degradation and the microtubule-associated protein 1B. (2016)
  • Author(s): Villarroel-Campos, D., Henriquez, D. R., Bodaleo, F. J., Oguchi, M. E., Bronfman, F. C., Fukuda, M. and Gonzalez-Billault, C.
  • Journal Title: J. Neurosci. Volume: 36 Issue: 27 Pages: 7298-7313
  • DOI: 10.1523/jneurosci.4064-15.2016
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Multiple roles of VARP in endosomal trafficking: Rabs, retromer components and R-SNARE VAMP7 meet on VARP. (2016)
  • Author(s): Fukuda, M.
  • Journal Title: Traffic Volume: 17 Issue: 7 Pages: 709-719
  • DOI: 10.1111/tra.12406
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Multiple types of guanine nucleotide exchange factors (GEFs) for Rab small GTPases. (2016)
  • Author(s): Ishida, M., Oguchi, M. E. and Fukuda, M.
  • Journal Title: Cell Struct. Funct. Volume: 41 Issue: 2 Pages: 61-79
  • DOI: 10.1247/csf.16008
  • NAID: 130005163247
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Regulation of podocalyxin trafficking by Rab small GTPases in epithelial cells. (2016)
  • Author(s): Mrozowska, P. S. and Fukuda, M.
  • Journal Title: Small GTPases Volume: 7 Issue: 4 Pages: 231-238
  • DOI: 10.1080/21541248.2016.1211068
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] RUTBC1 functions as a GTPase-activating protein for Rab32/38 and regulates melanogenic enzyme trafficking in melanocytes. (2016)
  • Author(s): Marubashi, S., Shimada, H., Fukuda, M. and Ohbayashi, N.
  • Journal Title: J. Biol. Chem. Volume: 291 Issue: 3 Pages: 1427-1440
  • DOI: 10.1074/jbc.m115.684043
  • NAID: 120007128493
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Rab12 regulates retrograde transport of mast cell secretory granules by interacting with the RILP-dynein complex. (2016)
  • Author(s): Efergan, A., Azouz, N. P., Klein, O., Noguchi, K., Rothenberg, M. E., Fukuda, M. and Sagi-Eisenberg, R.
  • Journal Title: J. Immunol. Volume: 196 Issue: 3 Pages: 1091-1101
  • DOI: 10.4049/jimmunol.1500731
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Small GTPase Rab2B and its specific binding protein Golgi-associated Rab2B interactor-like 4 (GARI-L4) regulate Golgi morphology. (2015)
  • Author(s): Aizawa, M. and Fukuda, M.
  • Journal Title: J. Biol. Chem. Volume: 290 Issue: 36 Pages: 22250-22261
  • DOI: 10.1074/jbc.m115.669242
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Measurement of Rab35 Activity with the GTP-Rab35 Trapper RBD35. (2015)
  • Author(s): Kobayashi H, Etoh K, Marubashi S, Ohbayashi N, Fukuda M.
  • Journal Title: Methods Mol Biol Volume: 1298 Pages: 207-216
  • DOI: 10.1007/978-1-4939-2569-8_18
  • ISBN: 9781493925681, 9781493925698
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Assay of Rab17 and its guanine nucleotide exchange factor Rabex-5 in the dendrites of hippocampal neurons. (2015)
  • Author(s): Mori, Y. and Fukuda, M.
  • Journal Title: Methods Mol. Biol. Volume: 1298 Pages: 233-243
  • DOI: 10.1007/978-1-4939-2569-8_20
  • ISBN: 9781493925681, 9781493925698
  • Peer Reviewed
• [Presentation] リサイクリングエンドソームに局在する新規 Rab11 結合タンパク質TBC1D12 は神経突起伸長を制御する (2017)
  • Author(s): 小口舞、野口憲太、福田光則
  • Organizer: 日本生化学会東北支部 第83回例会
• [Presentation] 神経突起伸長を負に制御する新規低分子量Gタンパク質Rabの探索 (2017)
  • Author(s): 小口舞、衛藤貫、福田光則
  • Organizer: 第69回日本細胞生物学会大会
• [Presentation] Rab small GTPases are key players for neurite outgrowth and differentiation. (2017)
  • Author(s): Mitsunori Fukuda
  • Organizer: Seminar in the National Center of Neurology and Psychiatry
  • Int'l Joint Research / Invited
• [Presentation] The small GTPase Rab10 regulates the formation of tubular endosomes. (2017)
  • Author(s): Kan Etoh and Mitsunori Fukuda
  • Organizer: The 2017 ASCB/EMBO meeting
  • Int'l Joint Research
• [Presentation] Rab8/10活性化因子・Rabin8による神経突起伸長の制御メカニズム (2016)
  • Author(s): 福田光則
  • Organizer: 第39回日本分子生物学会年会シンポジウム「メンブレントラフィック研究の新時代」
  • Place of Presentation: パシフィコ横浜（神奈川県、横浜市）
  • Year and Date: 2016-11-30
  • Invited
• [Presentation] リサイクリングエンドソームに局在する新規Rab11結合タンパク質TBC1D12の機能解析 (2016)
  • Author(s): 小口舞、野口憲太、福田光則
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター（宮城県・仙台市）
  • Year and Date: 2016-09-27
• [Presentation] 細胞外環境に依存したRabファミリーによる上皮細胞極性輸送の制御 (2016)
  • Author(s): 福田光則
  • Organizer: 第89回日本生化学会大会シンポジウム「膜動態を介した細胞間・細胞外環境との相互作用の制御」
  • Place of Presentation: 仙台国際センター（宮城県・仙台市）
  • Year and Date: 2016-09-25
  • Invited
• [Presentation] Regulation of podocalyxin trafficking by Rab small GTPases in epithelial cells. (2016)
  • Author(s): Mitsunori Fukuda
  • Organizer: 2016 FASEB Science Research Conference “GTPases in Trafficking, Autophagy and Disease”
  • Place of Presentation: West Palm Beach, FL (USA)
  • Year and Date: 2016-09-21
  • Int'l Joint Research / Invited
• [Presentation] Rab12 regulates podocalyxin exit from early endosomes through tubulation of endosomal membranes. (2016)
  • Author(s): Paulina S. Mrozowska and Mitsunori Fukuda
  • Organizer: 2016 FASEB Science Research Conference “GTPases in Trafficking, Autophagy and Disease”
  • Place of Presentation: West Palm Beach, FL (USA)
  • Year and Date: 2016-09-20
• [Presentation] リサイクリングエンドソームに局在する新規Rab11結合タンパク質TBC1D12の同定 (2016)
  • Author(s): 小口舞、野口憲太、福田光則
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ（京都府・京都市）
  • Year and Date: 2016-06-17
• [Presentation] Rab35活性化因子DENND1A/connecdenn 1は神経突起伸長を制御する (2016)
  • Author(s): 衛藤貫、福田光則
  • Organizer: 第68回日本細胞生物学会大会
  • Place of Presentation: 京都テルサ（京都府・京都市）
  • Year and Date: 2016-06-16
• [Presentation] 上皮細胞のポドカリキシン極性輸送におけるRabシグナリングネットワークの解析 (2016)
  • Author(s): 福田光則
  • Organizer: 第68回日本細胞生物学会大会シンポジウム「高次生命機能を司るメンブレントラフィック：分子基盤からその破綻による疾患発症の理解に向けて」
  • Place of Presentation: 京都テルサ（京都府・京都市）
  • Year and Date: 2016-06-15
  • Invited
• [Presentation] Regulation of podocalyxin trafficking by Rab small GTPases in 2D and 3D epithelial cell cultures. (2016)
  • Author(s): Paulina S. Mrozowska and Mitsunori Fukuda
  • Organizer: 2016 Gordon Research Conference “Cell Polarity and Signaling”
  • Place of Presentation: West Dover, VT (USA)
  • Year and Date: 2016-06-13
  • Int'l Joint Research
• [Presentation] 細胞内小胞輸送が制御する多彩な生命現象～メラニン輸送、オートファジー、ヒスタミン分泌まで～ (2016)
  • Author(s): 福田光則
  • Organizer: 第40回皮膚科免疫セミナー
  • Place of Presentation: 経団連会館（東京都、千代田区）
  • Year and Date: 2016-03-05
  • Invited
• [Presentation] 神経突起伸長におけるRabシグナリングカスケードの役割 (2015)
  • Author(s): 福田光則
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会ワークショップ「高次生命機能を支えるメンブレントラフィック」
  • Place of Presentation: 神戸ポートアイランド（兵庫県、神戸市）
  • Year and Date: 2015-12-03
  • Invited
• [Presentation] 神経突起伸長におけるRabin8の機能解析 (2015)
  • Author(s): 本間悠太、福田光則
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸ポートアイランド（兵庫県、神戸市）
  • Year and Date: 2015-12-01
• [Presentation] Rab35・centaurin-β2/ACAP2複合体の形成による神経突起伸長及び細胞質分裂の制御 (2015)
  • Author(s): 衛藤貫、福田光則
  • Organizer: 第67回日本細胞生物学会大会
  • Place of Presentation: タワーホール船堀（東京都、江戸川区）
  • Year and Date: 2015-06-30
• [Presentation] Rab small GTPases, key players in melanosome movements and neurite outgrowth. (2015)
  • Author(s): Mitsunori Fukuda
  • Organizer: 2015 Departmental Seminar Series at the Pasteur Institute
  • Place of Presentation: Paris, France
  • Year and Date: 2015-06-08
  • Int'l Joint Research / Invited
• [Book] Encyclopedia of Signaling Molecules 2nd Edition (Choi, S. ed.) "Slp (synaptotagmin-like protein) " (2018)
  • Author(s): Fukuda, M.
  • Publisher: Springer
  • ISBN: 9781493967995
• [Book] Encyclopedia of Signaling Molecules 2nd Edition (Choi, S. ed.) "Rab27" (2018)
  • Author(s): Oguchi, M. E. and Fukuda, M.
  • Publisher: Springer
  • ISBN: 9781493967995
• [Remarks] 東北大学大学院生命科学研究科膜輸送機構解析分野
  • URL: https://www.lifesci.tohoku.ac.jp/research/teacher/detail.html?id=1724
• [Remarks] 東北大学大学院生命科学研究科膜輸送機構解析分野
  • URL: http://www.lifesci.tohoku.ac.jp/teacher/t_fukuda/