Degradation mechanism of effector molecules by Rab small GTPase

• Project/Area Number: 15K07039
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Cell biology
• Research Institution: University of Tsukuba
• Principal Investigator: Ohbayashi Norihiko 筑波大学, 医学医療系, 准教授 (40421979)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2017: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2016: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 色素細胞 / Rab / プロテアソーム / メラニン合成酵素 / 低分子量Gタンパク質 / ユビキチン / 小胞輸送 / メラノソーム / 品質管理 / オルガネラ / タンパク質分解 / 蛋白質分解 / 低分子量G蛋白質

Outline of Final Research Achievements

Varp has been shown to function as a Rab32/Rab38 effector through its first ANKR1 domain. Although these functions of Varp are important for melanogenesis, Varp contains a second ANKR2 domain, whose function remained completely unknown. Here Rab40C was identified as a novel ANKR2-binding protein and it was investigated that Rab40C-involvement in melanogenic enzyme trafficking in melanocytes. The results here showed that overexpression of Rab40C in melanocytes caused a dramatic reduction in melanogenic enzyme Tyrp1 signals by promoting proteasomal degradation of Varp in a SOCS-box-dependent manner and that knockdown of Rab40C in melanocytes caused an increase in the amount of Varp. Intriguingly, Rab40C knockdown also caused a dramatic reduction in Tyrp1 signals, the same as Varp overexpression did. These findings indicated that Rab40C is a previously unexpected regulator of Tyrp1 trafficking in melanocytes through controlling the proteasomal degradation of Varp.

Research Products

• [Journal Article] The RAB2B-GARIL5 Complex Promotes Cytosolic DNA-Induced Innate Immune Responses (2017)
  • Author(s): Takahama Michihiro、Fukuda Mitsunori、Ohbayashi Norihiko、Kozaki Tatsuya、Misawa Takuma、Okamoto Toru、Matsuura Yoshiharu、Akira Shizuo、Saitoh Tatsuya
  • Journal Title: Cell Reports Volume: 20 Issue: 12 Pages: 2944-2954
  • DOI: 10.1016/j.celrep.2017.08.085
  • NAID: 120007134819
  • Peer Reviewed / Open Access
• [Journal Article] Arf6 in lymphatic endothelial cells regulates lymphangiogenesis by controlling directional cell migration (2017)
  • Author(s): Lin Yueh-Chien、Ohbayashi Norihiko、Hongu Tsunaki、Katagiri Naohiro、Funakoshi Yuji、Lee Hsinyu、Kanaho Yasunori
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 11431-11431
  • DOI: 10.1038/s41598-017-11240-x
  • NAID: 120007134837
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Regulation of HGF-induced hepatocyte proliferation by the small GTPase Arf6 through the PIP2-producing enzyme PIP5K1A (2017)
  • Author(s): Tsai Meng-Tsz、Katagiri Naohiro、Ohbayashi Norihiko、Iwasaki Kenichi、Ohkohchi Nobuhiro、Ding Shih-Torng、Kanaho Yasunori、Funakoshi Yuji
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 9438-9438
  • DOI: 10.1038/s41598-017-09633-z
  • NAID: 120007134880
  • 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] The small G protein Arf6 expressed in keratinocytes by HGF stimulation is a regulator for skin wound healing. (2017)
  • Author(s): Miura Y, Ngo Thai Bich V, Furuya M, Hasegawa H, Takahashi S, Katagiri N, Hongu T, Funakoshi Y, Ohbayashi N, Kanaho Y.
  • Journal Title: Scientific Reports Volume: 7 Issue: 1 Pages: 46649-46649
  • DOI: 10.1038/srep46649
  • NAID: 120007129142
  • Peer Reviewed / Open Access
• [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] ACAP3 regulates neurite outgrowth through its GAP activity specific to Arf6 in mouse hippocampal neurons. (2016)
  • Author(s): Miura Y, Hongu T, Yamauchi Y, Funakoshi Y, Katagiri N, Ohbayashi N, Kanaho Y.
  • Journal Title: Biochemical Journal Volume: 473 Issue: 17 Pages: 2591-2602
  • DOI: 10.1042/bcj20160183
  • Peer Reviewed / Open Access
• [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] Pathological functions of the small GTPase Arf6 in cancer progression: Tumor angiogenesis and metastasis (2016)
  • Author(s): Hongu T, Yamauchi Y, Funakoshi Y, Katagiri N, Ohbayashi N, Kanaho Y
  • Journal Title: Small GTPases Volume: 0 Issue: 2 Pages: 1-7
  • DOI: 10.1080/21541248.2016.1154640
  • Peer Reviewed / Open Access / Int'l Joint Research
• [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] Rab1A regulates anterograde melanosome transport by recruiting kinesin-1 to melanosomes through interaction with SKIP (2015)
  • Author(s): Ishida, M., Ohbayashi, N. and Fukuda, M
  • Journal Title: Sci. Rep. Volume: 5 Issue: 1 Pages: 8238-8238
  • DOI: 10.1038/srep08238
  • Peer Reviewed / Open Access
• [Journal Article] Rab40C is a novel Varp-binding protein that promotes proteasomal degradation of Varp in melanocytes. (2015)
  • Author(s): Yatsu, A., Shimada, H., Ohbayashi, N. and Fukuda, M.
  • Journal Title: Biol. Open Volume: 4 Issue: 3 Pages: 267-275
  • DOI: 10.1242/bio.201411114
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Activation of the Small G Protein Arf6 by Dynamin2 through Guanine Nucleotide Exchange Factors in Endocytosis. (2015)
  • Author(s): Okada R, Yamauchi Y, Hongu T, Funakoshi Y, Ohbayashi N, Hasegawa H, Kanaho Y
  • Journal Title: Sci Rep Volume: 5 Issue: 1 Pages: 14919-14919
  • DOI: 10.1038/srep14919
  • NAID: 120007135937
  • Peer Reviewed / Open Access / Int'l Joint Research / 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
• [Presentation] A Varp-binding protein, RACK1, regulates dendrite outgrowth through stabilization of Varp protein in melanocytes. (2017)
  • Author(s): Soujiro Marubashi, Norihiko Ohbayashi and Mitsunori Fukuda
  • Organizer: The 2017 Japan-NIH joint Symposium on Advances in Biomedical Research and Disease
  • Int'l Joint Research
• [Presentation] The molecular mechanisms of anterograde melanosome transport on microtubules in mammalian melanocytes. (2015)
  • Author(s): Ishida M, Ohbayashi N, Fukuda M
  • Organizer: The 2015 Annual Meeting of the American Society for Cell Biology
  • Place of Presentation: San Diego, CA
  • Year and Date: 2015-12-12
  • Int'l Joint Research
• [Presentation] RUTBC1はRab32/38の不活性化を通してメラニン合成酵素の輸送を調節する (2015)
  • Author(s): 大林典彦、丸橋総史郎、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: 神戸市、神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] 新規Varp結合分子・RACK1はVarpの安定化を介してメラノサイトのデンドライト伸長に関与する (2015)
  • Author(s): 丸橋総史郎、大林典彦、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: 神戸市、神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] 癌細胞浸潤における Arf6 GTPase 活性化因子 ARAP3 の機能解析 (2015)
  • Author(s): 山内庸平、 本宮綱記、山口英樹、船越祐司、大林典彦、金保安則
  • 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: 神戸市、神戸ポートアイランド
  • Year and Date: 2015-12-01
• [Presentation] The Arf6-specific GTPase-activating protein ARAP3 regulates breast cancer cell invasion. (2015)
  • Author(s): Yohei Y, Hongu T, Yamaguchi H, Funakoshi Y, Ohbayashi N, Kanaho Y
  • Organizer: Tsukuba Global Science Week 2015
  • Place of Presentation: Tsukuba, Japan
  • Year and Date: 2015-09-28
  • Int'l Joint Research
• [Presentation] ACAP3はArf6特異的なGAPとして働き、マウス海馬神経細胞の神経突起伸長を制御する (2015)
  • Author(s): 三浦悠樹、本宮綱記、船越祐司、大林典彦、金保安則
  • Organizer: 第14回生命科学研究会
  • Place of Presentation: 三浦市、三浦市民ホール
  • Year and Date: 2015-06-26
• [Presentation] Varpの新規結合分子の探索とメラノサイトのデンドライト形成への関与 (2015)
  • Author(s): 丸橋総史郎、大林典彦、福田光則
  • Organizer: 日本生化学会東北支部 第81回例会
  • Place of Presentation: 仙台市、東北大学片平さくらホール
  • Year and Date: 2015-05-09
• [Remarks]
  • URL: http://www.md.tsukuba.ac.jp/basic-med/biochem/kanaholab/
• [Remarks]
  • URL: http://www.md.tsukuba.ac.jp/basic-med/biochem/kanaholab/
• [Remarks]
  • URL: http://www.tohoku.ac.jp/japanese/newimg/pressimg/touhokuuniv-press_20150202_02web.pdf