Analysis of GBP receptor structure and GBP signal transduction in hemocytes

• Project/Area Number: 18370031
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Animal physiology/Animal behavior
• Research Institution: Saga University
• Principal Investigator: HAYAKAWA Yoichi Saga University, 農学部, 教授 (50164926)
• Co-Investigator(Kenkyū-buntansha): AIZAWA Tomoyasu 北海道大学, 大学院・理学研究科, 准教授 (40333596) ; 島田 公夫 北海道大学, 低温科学研究所, 助教 (30001663)
• Project Period (FY): 2006 – 2009
• Project Status: Completed (Fiscal Year 2009)
• Budget Amount: ¥18,060,000 (Direct Cost: ¥15,000,000、Indirect Cost: ¥3,060,000); Fiscal Year 2009: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2008: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2007: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2006: ¥4,800,000 (Direct Cost: ¥4,800,000)
• Keywords: 昆虫 / growth-blocking peptide / サイトカイン / GBP受容体アダプター / 血球細胞 / インテグリン / シグナル伝達 / チロシンリン酸化 / レセプター / GBP / 情報伝達 / GBPレセプター / 細胞質カルシウム / growth-blocking peptide(GBP) / 細胞膜タンパク質

Research Abstract

Growth-blocking peptide (GBP) is an insect cytokine that stimulates a class of immune cells called plasmatocytes to adhere to each other and a foreign surface. Although structure-activity studies have been extensively performed in GBP and its mutants of Lepidoptera Pseudaletia separata, the signaling pathway of GBP-dependent activation of plasmatocytes remains unknown. We identified a novel adaptor protein (P77) with a molecular mass of 77kDa containing SH2/SH3 domain binding motifs and an immunoreceptor tyrosine-based activation motif (ITAM)-like domain in the cytoplasmic region of the C-terminus. Although P77 did not have the capacity for direct binding with GBP, its cytoplasmic tyrosine residues were specifically phosphorylated within seconds after addition of GBP to a plasmatocyte suspension. Tyrosine phosphorylation of P77 was also observed when hemocytes were incubated with Enterobactor cloacae or Micrococcus luteus, but this phosphorylation was found to be induced by GBP released from hemocytes stimulated by the pathogens. We further detected tyrosine phosphorylation of the integrin β subunit in plasmatocytes stimulated by GBP. Double-stranded RNAs targeting P77 not only decreased GBP-dependent tyrosine phosphorylation of the integrin β subunit, but also abolished the GBP-induced spreading of plasmatocytes on foreign surfaces. P77 RNAi larvae also showed significantly higher mortality than control larvae following infection with Serratia marcescens, thus indicating that P77 is essential for GBP to mediate a normal innate cellular immunity in insects. These results demonstrated that GBP signaling in plasmatocytes requires the adaptor protein P77 and that active P77-assisted tyrosine phosphorylation of integrins is critical for the activation of plasmatocytes.

Research Products

• [Journal Article] The C-terminal elongation of Growth-blocking peptide enhances its biological activity and micelle binding affinity. (2009)
  • Author(s): Umetsu, Y, Aizawa, T, Muto, K, Yamamoto, H, Kamiya, M, Kumaki, Y, Demura, M, Hayakawa, Y., Kawano, K.
  • Journal Title: J. Biol. Chem. 284 Pages: 29625-29634
• [Journal Article] A novel peptide mediates aggregation and migration of hemocytes from an insect. (2009)
  • Author(s): Nakatogawa, S., Oda, Y., Kamiya, M., Kamijima, T., Aizawa, T., Clark, KD., Kawano, K., Strand, R.M., Hayakawa, Y.
  • Journal Title: Curr. Biol. 19 Pages: 779-785
• [Journal Article] The C-terminal elongation of Growth-blocking peptide enhances its biological activity and micelle binding affinity. (2009)
  • Author(s): Umetsu, Y, Aizawa, T, Muto, K, Yamamoto, H, Kamiya, M, Kumaki, Y, Demura, M, Hayakawa, Y., Kawano, K.
  • Journal Title: J.Biol.Chem. 284 Pages: 29625-29634
  • Peer Reviewed
• [Journal Article] A novel peptide mediates aggregation and migration of hemocytes from an insect. (2009)
  • Author(s): Nakatogawa, S., Oda, Y., Kamiya, M., Kamijima, T., Aizawa, T., Clark, KD., Kawano, K., Strand, R.M., Hayakawa, Y.
  • Journal Title: Curr.Biol. 19 Pages: 779-785
  • Peer Reviewed
• [Journal Article] A novel peptide mediates aggregation and migration of hemocytes from an insect (2009)
  • Author(s): Nakatogawa, S., Oda, Y., Kamiya, M., Aizawa, T., Kawano,K., Strand, R.M. and Hayakawa, Y.
  • Journal Title: Curr. Biol. (In press)
• [Journal Article] A novel peptide mediates aggregation and migration ofhemocytes from an insect. (2009)
  • Author(s): Nakatogawa, S.
  • Journal Title: Curr. Biol. (In press)
  • Peer Reviewed
• [Journal Article] A gene involved in the food preferences of larval Drosophila melanogaster. (2008)
  • Author(s): Ryuda, M., Tsuzuki, S., Tanimura, T., Tojo, S., Hayakawa, Y.
  • Journal Title: J. Insect Physiol. 54 Pages: 1440-1445
• [Journal Article] Strains in the common cutworm, Spodoptera litura (Lepidoptera: Noctuidae) with differing host ranges. (2008)
  • Author(s): Tojo, S., Hayakawa, Y., Phaophan P.
  • Journal Title: 43 Pages: 491-496
  • NAID: 110007004670
• [Journal Article] A novel gene associated with intraspecific predation in Spodoptera litura larvae. (2008)
  • Author(s): Ryuda, M., Nakayama, H., Hayakawa, Y.
  • Journal Title: Appl. Entomol. Zool. 43 Pages: 563-568
  • NAID: 110007004680
• [Journal Article] Analysis of hunger-driven gene expression in Drosophila melanogaster larval central nervous system. (2008)
  • Author(s): Ryuda, M., Shimada, K., Koyanagi, R., Azumi, K., Tanimura, T., Hayakawa, Y.
  • Journal Title: Zool. Sci. 25 Pages: 746-752
  • NAID: 40016249920
• [Journal Article] Insect cytokine growth-blocking peptide signaling cascades regulate two separate groups of target genes. (2008)
  • Author(s): Ninomiya, Y., Kurakake, M., Oda, Y., Tsuzuki, S., Hayakawa, Y.
  • Journal Title: FEBS J. 275 Pages: 894-902
• [Journal Article] Drosophila melanogaster larvalcentral nervous system (2008)
  • Author(s): Ryuda, M.,Shimada, K., Koyanagi, R. Tanimura, T. and Hayakawa, Y.
  • Journal Title: Zool.Sci. 25 Pages: 746-75
• [Journal Article] A novel gene associated with intraspecific predation inSpodoptera litura larvae (2008)
  • Author(s): Ryuda, M., Nakayama, H. and Hayakawa, Y.
  • Journal Title: Appl. Entomol. Zool. 43 Pages: 563-568
• [Journal Article] Strains in the common cutworm Spodoptera litura (2008)
  • Author(s): Tojo, S., Hayakawa, Y., Phaophan P.
  • Journal Title: (Lepidoptera: Noctuidae) with differing host ranges. Pages: 491-496
• [Journal Article] A gene involved in the food preferences of larval Drosophila melanogaster (2008)
  • Author(s): Ryuda, M., Tsuzuki, S., Tanimura, T., Tojo, S. and Hayakawa, Y.
  • Journal Title: J. Insect Physiol. 54 Pages: 1440-1445
• [Journal Article] Analysis of hunger-driven gene expression in Drosophilamelanogaster larval central nervous system. (2008)
  • Author(s): Ryuda, M.
  • Journal Title: Z001. Sci. 25 Pages: 746-752
  • Peer Reviewed
• [Journal Article] A novel gene associated with intraspecific predation in Spodopteralitura larvae. (2008)
  • Author(s): Ryuda, M.
  • Journal Title: Appl. Entomol. Zool. 43 Pages: 563-568
  • Peer Reviewed
• [Journal Article] Strains in the common cutworm, Spodoptera litura (Lepidoptera : Noctuidae) with differing host ranges. (2008)
  • Author(s): Tojo, S.
  • Journal Title: Appl. Entomol. Zool. 43 Pages: 491-496
  • Peer Reviewed
• [Journal Article] A gene involved in the food preferences of larval Drosophilamelanogaster. (2008)
  • Author(s): Ryuda, M.
  • Journal Title: J. Insect Physiol. 54 Pages: 1440-1445
  • Peer Reviewed
• [Journal Article] Insect cytokine growth-blocking peptide signaling cascades regulate two separate groups of target genes. (2008)
  • Author(s): Ninomiya, Y., Kurakake, M., Oda, Y., Tsuzuki, S.and Hayakawa, Y.
  • Journal Title: FEBS Journal 275 Pages: 894-902
• [Journal Article] ようやく始まった昆虫サイトカイン研究 (2008)
  • Author(s): 早川 洋一
  • Journal Title: 化学と生物 46 Pages: 274-280
  • NAID: 10021177174
• [Journal Article] Insect cytokine, growth-blocking peptide, is a primary regulator of melanin-synthesis enzymes in armyworm larval cuticle. (2007)
  • Author(s): Ninomiya, Y., Hayakawa, Y.
  • Journal Title: FEBS J. 274 Pages: 1768-1777
• [Journal Article] Insect cytokine,growth-blocking peptide,is a primary regulator of melanin-synthesis enzymes in armyworm larval cuticle. (2007)
  • Author(s): Ninomiya, Y.and Hayakawa, Y.
  • Journal Title: FEBS Journal 274 Pages: 638-645
• [Journal Article] Insect cytokine, growth-blocking peptide, is a primary regulator of melanin-synthesis enzymes in armyworm larval cuticle. (2007)
  • Author(s): Ninomiya, Y., Hayakawa, Y.
  • Journal Title: FEBS Journal (In press)
• [Journal Article] Insect cytokine growth-blocking peptide (GBP) regulates insect development. (2006)
  • Author(s): Hayakawa, Y.
  • Journal Title: Appl. Entomol. Zool. 41 Pages: 545-554
  • NAID: 110006148628
• [Journal Article] N-Terminal Mutational Analysis of the Interaction Between Growth-Blocking Peptide (GBP) and Receptor of Insect Immune Cells. (2006)
  • Author(s): Watanabe, S., Tada, M., Aizawa, T., Yoshida, M., Sugaya, T., Taguchi, M., Kouno, T., Nakamura, T., Mizuguchi, M., Demura, M. Hayakawa, Y., Kawano, K.
  • Journal Title: Protein & Peptide Letters 13 Pages: 815-822
• [Journal Article] Mechanisms of black and white stripe pattern formation in the cuticles of insect larvae. (2006)
  • Author(s): Ninomiya, Y., Tanaka, K., Hayakawa, Y.
  • Journal Title: J. Insect Physiol. 52 Pages: 638-645
• [Journal Article] Mechanisms o black and white stripe pattern formation in the cuticles of insect larvae. (2006)
  • Author(s): Ninomiya, Y., Tanaka, K., Hayakawa, Y.
  • Journal Title: J. Insect Physiol. 52 Pages: 638-645
• [Journal Article] N-Terminal Mutational Analysis of the Interaction Between Growth-Blocking Peptide (GBP) and Receptor of Insect Immune Cells. (2006)
  • Author(s): Watanabe, S., Tada, M., Aizawa, T., Yoshida, M., Sugaya, T., Taguchi, M., Kouno, T., Nakamura, T., Mizuguchi, M., Demura, M., Hayakawa, Y., Kawano, K.
  • Journal Title: Protein ＆ Peptide Letters 13 Pages: 815-822
• [Presentation] 寄生蜂の毒液が宿主ショウジョウバエに及ぼす致死作用 (2010)
  • Author(s): 降幡駿介、早川洋一
  • Organizer: 日本応用動物昆虫学会
  • Place of Presentation: 千葉
  • Year and Date: 2010-03-26
• [Presentation] 寄生蜂Asobara japonicaの毒液が宿主ショウジョウバエに及ぼす致死作用 (2010)
  • Author(s): 降幡駿介、早川洋一
  • Organizer: 応用動物昆虫学会
  • Place of Presentation: 千葉大学
  • Year and Date: 2010-03-26
• [Presentation] 新規サイトカインの構造と生理機能 (2009)
  • Author(s): 早川洋一
  • Organizer: 日本生化学会
  • Place of Presentation: 神戸
  • Year and Date: 2009-10-28
• [Presentation] 新規昆虫サイトカインの構造と生理機能 (2009)
  • Author(s): 早川洋一
  • Organizer: 生化学会
  • Place of Presentation: 神戸国際会議場
  • Year and Date: 2009-10-21
• [Presentation] Insect cytokinesregulating innate immune defences (2009)
  • Author(s): Yoichi, Hayakawa
  • Organizer: 日本農芸化学会 2009 年大会
  • Year and Date: 2009-03-29
• [Presentation] 新規昆虫サイトカインの構造と生理機能 (2009)
  • Author(s): 早川洋一
  • Organizer: 日本農芸化学会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2009-03-29
• [Presentation] 昆虫サイトカインの構造と生理機能 (2009)
  • Author(s): 早川洋一
  • Organizer: 日本農芸化学会
  • Place of Presentation: 福岡
  • Year and Date: 2009-03-28
• [Presentation] Insect cytokine regulating innate immune defences. (2008)
  • Author(s): 早川洋一
  • Organizer: 生物物理学会
  • Place of Presentation: 福岡
  • Year and Date: 2008-12-20
• [Presentation] 新規昆虫サイトカインの構造と生理機能 (2008)
  • Author(s): 早川, 洋一
  • Organizer: 日本農芸化学会 2009 年大会
  • Year and Date: 2008-12-04
• [Presentation] Characterization of a novel insect cytokine : itsStructure and functions (2008)
  • Author(s): 早川洋一
  • Organizer: 日本生物物理学会
  • Place of Presentation: 福岡国際会議場
  • Year and Date: 2008-12-04
• [Presentation] 細菌感染によるP77のリン酸化 (2008)
  • Author(s): 松本均、早川洋一
  • Organizer: 日本動物学会
  • Place of Presentation: 福岡
  • Year and Date: 2008-09-28
• [Presentation] コクヌストモドキの擬死行動誘発に関連する遺伝子解析 (2008)
  • Author(s): 田崎彩子、早川洋一
  • Organizer: 日本動物学会
  • Place of Presentation: 福岡
  • Year and Date: 2008-09-28
• [Presentation] イロショウジョウバエの食性・摂食行動に関わるCG33071遺伝子の解析 (2008)
  • Author(s): 龍田勝輔、早川洋一
  • Organizer: 日本動物学会
  • Place of Presentation: 福岡
  • Year and Date: 2008-09-28
• [Presentation] コクヌストモドキの擬死行動誘発に関連する遺伝子解析 (2008)
  • Author(s): 早川洋一
  • Organizer: 日本動物学会
  • Place of Presentation: 福岡大学
  • Year and Date: 2008-09-06
• [Presentation] コクヌストモドキの擬死行動誘発に関連する遺伝子の解析 (2008)
  • Author(s): 田崎彩子、酒井美幸、宮竹貴久、早川洋一
  • Organizer: 日本動物学会第79回大会
• [Presentation] 細菌感染によるP77のリン酸化 (2008)
  • Author(s): 松本均、織田康則、早川洋一
  • Organizer: 日本動物学会第79回大会
• [Presentation] キイロショウジョウバエの食性・摂食行動に関わるCG33071遺伝子の解析 (2008)
  • Author(s): 龍田勝輔、谷村禎一、早川洋一
  • Organizer: 日本生物物理学会 第46回年会
• [Presentation] 昆虫の食性決定に関与する遺伝子の解析 (2007)
  • Author(s): 龍田勝輔、早川洋一
  • Organizer: 日本応用動物昆虫学会
  • Place of Presentation: 広島
  • Year and Date: 2007-03-28
• [Presentation] ホームページ等
  • Author(s): http://extwww. cc. saga-u. ac. jp/~hayakayo /FRAME/index. html
• [Remarks]
  • URL: http://extwww.cc.saga-u.ac.jp/~hayakayo/FRAME/