メラノソーム輸送におけるRab27Aエフェクターのヒエラルキーの解析

• Project/Area Number: 16044248
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: 福田 光則 独立行政法人理化学研究所, 福田独立主幹研究ユニット, 独立主幹研究員 (50311361)
• Project Period (FY): 2004 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥6,500,000 (Direct Cost: ¥6,500,000); Fiscal Year 2005: ¥3,200,000 (Direct Cost: ¥3,200,000); Fiscal Year 2004: ¥3,300,000 (Direct Cost: ¥3,300,000)
• Keywords: シナプトタグミン様蛋白質 / Rab27A / Rabエフェクター / メラノソーム輸送 / Griscelli症候群 / ミオシンVa / Slac2-a / melanophilin / 膜輸送 / エキソサイトーシス / melanophili

Research Abstract

低分子量G蛋白質Rab27Aを欠損するヒト遺伝病タイプII型Griscelli症候群の患者では、乳幼児でも白髪様の毛髪を示すことが知られている。Griscelli症候群患者のメラノサイト(メラニン色素産生細胞>ではメラノソーム(メラニン色素含有小胞)の形成・成熟ではなく、メラノソームの輸送過程に異常が認められるが、その分子メカニズムはこれまで明らかではなかった。本研究では、Rab27Aに特異的に結合するエフェクター分子群に焦点を当て、Rab27Aを介するメラノソーム輸送の分子メカニズムの全貌を解明することに成功した(Nature Cell Biol.(2004)6, 1195-1203)。すなわち、メラノサイトにおいては二種類のRab27Aエフェクター(Slac2-a及びSlp2-a)が豊富に発現しており、Rab27Aはこれらの分子を連続的且つ秩序立って用いることによりメラノソームを核周辺から細胞膜まで輸送していたのである。まず、第一のエフェクターSlac2-aがメラノソーム上のRab27Aとアクチン依存性モーター蛋白質であるミオシンVaと三者複合体を形成し、メラノソームを核周辺から細胞膜周辺へと輸送し、次いで第二のエフェクターSlp2-aと結合することによりメラノソームを細胞膜へつなぎ止める(Slp2-aのC2Aドメインが細胞膜の構成成分であるホスファチジルセリンと結合する)ことが明らかとなった。また、Slac2-aとSlp2-aのRab27A結合能を比較してみると、Slp2-aの方がRab27Aに高親和性に結合し、解離速度も遅いことからメラノソームの細胞膜へのつなぎ止めに適していることも明らかとなった。

Research Products

• [Journal Article] Distinct developmental expression of synaptotagmin I and IX in the mouse brain. (2006)
  • Author(s): Fukuda, M.
  • Journal Title: NeuroReport 17 Pages: 179-182
• [Journal Article] O-glycosylation is essential for intracellular targeting of synaptotagmins I and II in non-neuronal specialized secretory cells. (2005)
  • Author(s): Atiya-Nasagi, Y., Cohen, H., Medalia, O., Fukuda, M., Sagi-Eisenberg, R.
  • Journal Title: J.Cell Sci. 118 Pages: 1363-1372
• [Journal Article] Classical protein kinase C(s) regulates targeting of synaptotagmin IX to the endocytic recycling compartment. (2005)
  • Author(s): Haberman, Y., Ziv, I., Gorzalczany, Y., Fukuda, M., Sagi-Eisenberg, R.
  • Journal Title: J.Cell Sci. 118 Pages: 1641-1649
• [Journal Article] Functional analysis of Slac2-c/MyRIP as a linker protein between melanosomes and myosin VIIa. (2005)
  • Author(s): Kuroda, T.S., Fukuda, M.
  • Journal Title: J.Biol.Chem. 280 Pages: 28015-28022
• [Journal Article] AS160, the Akt substrate regulating GLUT4 translocation, has a functional Rab GTPase activating protein domain. (2005)
  • Author(s): Miinea, C.P., Sano, H., Kane, S., Sano, E., Fukuda, M., Peranen, J., Lane, W.S., Lienhard, G.E.
  • Journal Title: Biochem.J. 391 Pages: 87-93
• [Journal Article] Protein Kinase C-mediated phosphorylation of the two polybasic regions of synaptotagmin VI regulates their function in acrosomal exocytosis. (2005)
  • Author(s): Roggero, C.M., Tomes, C.N., De Blas, G.A., Castillo, J., Michaut, M.A., Fukuda, M., Mayorga, L.S.
  • Journal Title: Dev.Biol. 285 Pages: 422-435
• [Journal Article] Adenovirus-mediated silencing of Synaptotagmin 9 inhibits Ca^<2+>-dependent insulin secretion in islets. (2005)
  • Author(s): Iezzi, M., Eliasson, L., Fukuda, M., Wollheim, C.B.
  • Journal Title: FEBS Lett. 579 Pages: 5241-5246
• [Journal Article] Rab proteins regulate epithelial sodium channel activity in colonic epithelial HT-29 cells. (2005)
  • Author(s): Saxena, S., Singh, M., Engisch, K., Fukuda, M., Kaur, S.
  • Journal Title: Biochem.Biophys.Res.Commun. 337 Pages: 1219-1223
• [Journal Article] The short apical membrane half-life of rescued ΔF508-cystic fibrosis transmembrane conductance regulator(CFTR)CFTR results from accelerated endocytosis of ΔF508-CFTR in polarized human airway epithelial cells. (2005)
  • Author(s): Swiatecka-Urban, A., Brown, A., Moreau-Marquis, S., Renuka, J., Coutermarsh, B., Barnaby, R., Karlson, K.H., Flotte, T.R., Fukuda, M., Langford, G.M., Stanton, B.A.
  • Journal Title: J.Biol.Chem. 280 Pages: 36762-36772
• [Journal Article] Slp4-a/granuphilin-a interacts with syntaxin-2/3 in a Munc18-2-dependent manner. (2005)
  • Author(s): Fukuda, M., Imai, A., Nashida, T., Shimomura, H.
  • Journal Title: J.Biol.Chem. 280 Pages: 39175-39184
• [Journal Article] The C2B domain of rabphilin directly interacts with SNAP-25 and regulates the docking step of dense core vesicle exocytosis in PC12 cells. (2005)
  • Author(s): Tsuboi, T., Fukuda, M.
  • Journal Title: J.Biol.Chem. 280 Pages: 39253-39259
• [Journal Article] Cloning and genomic characterization of sytdep, a new synaptotagmin XIV-related gene. (2005)
  • Author(s): Herrero-Turrion, M.J., Fukuda, M., Mollinedo, F.
  • Journal Title: Biochem.Biophys.Res.Commun. 340 Pages: 386-394
• [Journal Article] Slp homology domain : A novel protein motif that specifically binds small GTPase Rab27. (2005)
  • Author(s): Fukuda, M.
  • Journal Title: Recent Res.Dev.Biochem. 6 Pages: 13-29
• [Journal Article] Versatile role of Rab27 in membrane trafficking : Focus on the Rab27 effector families. (2005)
  • Author(s): Fukuda, M.
  • Journal Title: J.Biochem. 137 Pages: 9-16
  • NAID: 10017342344
• [Journal Article] Functional analysis of Slac2-a/melaonophilin as a linker protein between Rab27A and myosin Va in melanosome transport. (2005)
  • Author(s): Kuroda, T.S., Fukuda, M.
  • Journal Title: Methods Enzymol. 403 Pages: 419-431
• [Journal Article] Identification and biochemical analysis of Slac2-c/MyRIP as a Rab27-, myosin Va/VIIa-, and actin-binding protein. (2005)
  • Author(s): Kuroda, T.S., Fukuda, M.
  • Journal Title: Methods Enzymol. 403 Pages: 431-444
• [Journal Article] Analysis of the role of Rab27 effector Slp4-a/granuphilin-a in dense-core vesicle exocytosis. (2005)
  • Author(s): Fukuda, M., Kanno, E.
  • Journal Title: Methods Enzymol. 403 Pages: 445-457
• [Journal Article] Assay and functional interactions of Rim2 with Rab3. (2005)
  • Author(s): Fukuda, M.
  • Journal Title: Methods Enzymol. 403 Pages: 457-468
• [Journal Article] Assay of Rab binding specificity of rabphilin and Noc2 : Target molecules for Rab27. (2005)
  • Author(s): Fukuda, M., Yamamoto, A.
  • Journal Title: Methods Enzymol. 403 Pages: 469-481
• [Journal Article] Slac2-aとSlp2-a:メラニン色素輸送のしくみ (2005)
  • Author(s): 福田光則
  • Journal Title: 医学のあゆみ 212 Pages: 769-770
• [Journal Article] シナプトタグミンと類似タンパク質による膜輸送制御の分子メカニズム (2005)
  • Author(s): 福田光則
  • Journal Title: 生化学 77 Pages: 603-618
  • NAID: 10016603083
• [Journal Article] 「メラニン色素」輸送の分子メカニズム:肌や髪の毛が黒くなる仕組み (2005)
  • Author(s): 福田光則
  • Journal Title: 日皮協ジャーナル 54 Pages: 60-66
• [Journal Article] 美白への新たなアプローチ-メラニン輸送をストップさせる- (2005)
  • Author(s): 黒田垂歩, 福田光則
  • Journal Title: BIONICS 2(11) Pages: 68-71
  • NAID: 40007003528
• [Journal Article] Muncl3-4 is a GTP-Rab27-binding protein regulating dense core granule serection in platelets. (2004)
  • Author(s): Shirakawa, R., Higashi, T., Tabuchi, A., Yoshioka, A., Nishioka, H., Fukuda, M., Kita, T., Horiuchi.H.
  • Journal Title: J.Biol.Chem. 279 Pages: 10730-10737
• [Journal Article] Synaptotagmin interaction with the syntaxin/SNAP-25 dimer is mediated by an evolutionarily conserved motif and is sensitive to inositol hexakisphosphate. (2004)
  • Author(s): Rickman, C., Archer, D.A., Meunier, F.A., Craxton, M., Fukuda, M., Burgoyne, R.D., Davletov, B.
  • Journal Title: J.Biol.Chem. 279 Pages: 12574-12579
• [Journal Article] Rabphilin and Noc2 are recruited to dense-core vesicles through specific interaction with Rab27A in PC12 cells. (2004)
  • Author(s): Fukuda, M., Kanno, E., Yamamoto, A.
  • Journal Title: J.Biol.Chem. 279 Pages: 13065-13075
• [Journal Article] The small GTPase Rab27B regulates amylase release from rat parotid acinar cells. (2004)
  • Author(s): Imai, A., Yoshie, S., Nashida, T., Shimomura, H., Fukuda, M.
  • Journal Title: J. Cell Sci. 117 Pages: 1945-1953
• [Journal Article] Slac2-a/melanophilin contains multiple PEST-like sequences that are highly sensitive to proteolysis. (2004)
  • Author(s): Fukuda, M., Itoh, T.
  • Journal Title: J.Biol.Chem. 279 Pages: 22314-22321
• [Journal Article] RNA interference-mediated silencing of synaptotagmin IX, but not synaptotagmin I, inhibits dense-core vesicle exocytosis in PC12 cells. (2004)
  • Author(s): Fukuda, M.
  • Journal Title: Biochem.J. 380 Pages: 875-879
• [Journal Article] Synaptotagmin V and IX isoforms control Ca^<2+>-dependent insulin exocytosis. (2004)
  • Author(s): Iezzi, M., Kouri, G., Fukuda, M., Wollheim, C.B.
  • Journal Title: J. Cell Sci. 117 Pages: 3119-3127
• [Journal Article] Synaptotagmin IV regulates glial glutamate release. (2004)
  • Author(s): Zhang, Q., Fukuda, M., Van Bockstaele, E., Pascual, O., Haydon, P.G.
  • Journal Title: Proc.Natl.Acad.Sci. USA 101 Pages: 9441-9446
• [Journal Article] Alternative splicing in the first α-helical region of the Rab-binding domain of Rim regulates Rab3A binding activity : Is Rim a Rab3 effector protein during evolution? (2004)
  • Author(s): Fukuda, M.
  • Journal Title: Genes Cells 9 Pages: 831-842
• [Journal Article] Effect of forskolin on synaptotagmin IV protein trafficking in PC12 cells. (2004)
  • Author(s): Fukuda, M., Yamamoto, A.
  • Journal Title: J.Biochem. 136 Pages: 245-253
  • NAID: 10016201637
• [Journal Article] Synaptotagmin VII is targeted to dense-core vesicles and regulates their Ca^<2+>-dependent exocytosis in PC12 cells. (2004)
  • Author(s): Fukuda, M., Kanno, E., Satoh, M., Saegusa, C., Yamamoto, A.
  • Journal Title: J.Biol.Chem. 279 Pages: 52677-52684
• [Journal Article] Rab27A-binding protein Slp2-a is required for peripheral melanosome distribution and elongated cell shape in melanocytes. (2004)
  • Author(s): Kuroda, T.S., Fukuda, M.
  • Journal Title: Nature Cell Biol. 6 Pages: 1195-1203
• [Journal Article] Vesicular reuptake inhibition by a synaptotagmin I C2B domain antibody at the squid giant synapse. (2004)
  • Author(s): Llinas, R.R., Sugimori, M., Moran, K., Moreira, J.E., Fukuda, M.
  • Journal Title: Proc.Natl.Acad.Sci. USA 101 Pages: 17855-17860
• [Journal Article] Rabphilin and Noc2 function as Rab27 effectors that control Ca^<2+>-regulated exocytosis. (2004)
  • Author(s): Fukuda, M.
  • Journal Title: Recent Res.Dev.Neurochem. 7 Pages: 57-69
• [Journal Article] シナプトタグミンによる調節性分泌の制御 (2004)
  • Author(s): 福田光則
  • Journal Title: 蛋白質 核酸 酵素 49 Pages: 2186-2197
  • NAID: 40006462984
• [Book] アンチ・エイジングシリーズ1「白髪・脱毛・育毛の実際」メラニン色素輸送メカニズムの解明 (2005)
  • Author(s): 福田光則, 黒田垂歩
  • Publisher: エヌ・ティー・エス