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Analysis of molecular mechanisms for apoptosis in insects infected with baculoviruses

Research Project

Project/Area Number 14206007
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field 蚕糸・昆虫利用学
Research InstitutionNagoya University

Principal Investigator

KOBAYASHI Michihiro  Nagoya University, Graduate School of Bioagricultural Sciences, Professor, 大学院・生命農学研究科, 教授 (60111837)

Co-Investigator(Kenkyū-buntansha) IKEDA Motoko  Nagoya University, Graduate School of Bioagricultural Sciences, Associate Professor, 大学院・生命農学研究科, 助教授 (20262892)
NIIMI Teruyuki  Nagoya University, Graduate School of Bioagricultural Sciences, Assistant Professor, 大学院・生命農学研究科, 助手 (00293712)
河村 敏  岐阜県生物技術研究所, 専門研究員
Project Period (FY) 2002 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥53,560,000 (Direct Cost: ¥41,200,000、Indirect Cost: ¥12,360,000)
Fiscal Year 2005: ¥6,370,000 (Direct Cost: ¥4,900,000、Indirect Cost: ¥1,470,000)
Fiscal Year 2004: ¥5,980,000 (Direct Cost: ¥4,600,000、Indirect Cost: ¥1,380,000)
Fiscal Year 2003: ¥13,390,000 (Direct Cost: ¥10,300,000、Indirect Cost: ¥3,090,000)
Fiscal Year 2002: ¥27,820,000 (Direct Cost: ¥21,400,000、Indirect Cost: ¥6,420,000)
Keywordsnucleopolyhedrovirus / Ld652Y cell / fall webworm / gypsy moth / apoptosis / global protein synthesis shutdown / anti-viral responce / バキュロウイルス / 組換えウイルス / gp64 / 培養細胞 / sf9細胞 / High Five細胞 / 昆虫
Research Abstract

Ld652Y cells were infected with seven different nucleopolyhedroviruses (NPVs), including those from Autographa californica (AcMNPV), Bombyx mori (BmNPV), Hyphantria cunea (HycuNPV), Spodoptera exigua (SeMNPV), Lymantria dispar, Orgyia pseudotsugata (OpMNPV) and Spodoptera litura (SpltMNPV). The results showed that Ld652Y cells infected with BmNPV, HycuNPV, SeMNPV, OpMNPV and SpltMNPV underwent apoptosis. In HycuNPV-infected Ld652Y cells, a considerable amount of viral DNA was synthesized although there was no detectable yield of budded virions and polyhedrin. Northern blot and immunoblot analyses revealed that HycuNPV inhibitor of apoptosis 3 (Hycu-IAP3), which has been shown to function in Sf9 cells, was expressed in HycuNPV-infected Ld652Y cells at a level comparable with that in HycuNPV-infected SpIm cells, which produced a high titer of progeny virions without any apoptotic response. These results imply that the relative ease of apoptosis induction in NPV-infected Ld652Y cells is l … More argely dependent on inherent cellular properties rather than functions of the respective NPVs, and indicate that the defect in progeny virion production is not merely due to the virus-induced apoptosis in HycuNPV-infected Ld652Y cells.
HycuNPV infection protected SpIm cells from actinomycin D-induced apoptosis as early as 4 h postinfection. Analysis by Southern hybridization revealed that the HycuNPV genome possessed three members of iap genes that were designated as hycu-iap1,hycu-iap2 and hycu-iap3 because of their amino acid sequence homology with iaps identified in other baculoviruses. Functional analysis of Hycu-IAPs by transient expression assay in Sf9 cells revealed that Hycu-IAP3 blocked apoptosis induced by actinomycin D and rescued replication of p35-deficient-mutant AcMNPV, while Hycu-IAP1 and Hycu-IAP2 did not show any anti-apoptotic functions. Knockdown of hycu-iap3 expression by RNAi during HycuNPV infection of permissive SpIm cells induced apoptosis. These results indicate that Hycu-IAP3 is essential for blockage of apoptosis during HycuNPV infection of permissive SpIm cells. Less

Report

(5 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (33 results)

All 2006 2005 2004 Other

All Journal Article (22 results) Publications (11 results)

  • [Journal Article] A recombinant Bombyx mori nucleopolyhyedrovirus possessing hrf-1 gene replicates in nonpermissive Lymantria dispar IPLB-652Y cell line2006

    • Author(s)
      Ishikawa, H., Ogasawara, T., Ikeda, M., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 75・1

      Pages: 31-38

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Abortive replication of Bombyx mori nucleopolyhyedrovirus in Sf9 and High Five cells : Defective nuclear transport of the virions2006

    • Author(s)
      Katou, Y., Ikeda, M., Kobayashi, M.
    • Journal Title

      Virology 347・2

      Pages: 455-465

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] hycu-hr6, a large homologous region of Hyphantria cunes nucleopolyhedrovirus, is dispensable for virus replication in cell culture2006

    • Author(s)
      Nagai, S., Felipe Alves, C.A., Shirata, N., Katou, Y., Ishikawa, H., Ikeda, M., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 75・2(in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome2006

    • Author(s)
      Ikeda, M., Shikata, N., Shirata, N., Chaeychomsri, S., Kobayashi, M.
    • Journal Title

      Journal of General Virology 87(in press)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A recombinant Bombyx mori nucleopolyhedrovirus possessing hrf-1 gene replicates in nonpermissive Lymantria dispar IPLB-Ld652Y cell line.2006

    • Author(s)
      Ishikawa, H., Ogasawara, T., Ikeda, M., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 75(1)

      Pages: 31-38

    • NAID

      10017316184

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Abortive replication of Bombyx mori nucleopolyhedrovirus in Sf9 and High Five cells : Defective nuclear transport of the virions.2006

    • Author(s)
      Katou, Y., Ikeda, M., Kobayashi, M.
    • Journal Title

      Virology 347(2)

      Pages: 455-465

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] hycu-hr6, a large homologous region of Hyphantria cunea nucleopolyhedrovirus, is dispensable for virus replication in cell culture.2006

    • Author(s)
      Nagai, S., Felipe Alves, C.A., Shirata, N., Katou, Y., Ishikawa, H., Ikeda, M., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 75(2) (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome.2006

    • Author(s)
      Ikeda, M., Shikata, M., Shirata, N., Chaeychomsri, S., Kobayashi, M.
    • Journal Title

      Journal of General Virology 87 (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] A recombinant Bombyx mori nucleopolyhedrovirus possessing hrf-1 gene replicates in nonpermissive Lymantria dispar IPLB-Ld652Y cell line2006

    • Author(s)
      Ishikawa, H., Ogasawara, T., Ikeda, M., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 75・1

      Pages: 31-38

    • NAID

      10017316184

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Abortive replication of Bombyx mori nucleopolyhedrovirus in Sf9 and High Five cells : Defective nuclear transport on the virions2006

    • Author(s)
      Katou, Y., Ikeda, M., Kobayashi, M.
    • Journal Title

      Virology (in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] hycu-hr, a large homologous region of Hyphantria cunea nucleopolyhedrovirus, is dispensable for virus replication in cell culture2006

    • Author(s)
      Nagai, S., Felipe Alves, C.A., Shirata, N., Katou, Y., Ishikawa, H., Ikeda, M., Kobayashi, M..
    • Journal Title

      Journal of Insect Biotechnology and Sericology 75・2(in press)

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Accumulation of non-viral 45.5K and 44K polypeptides inversely correlates with that of viral structural polypeptides in the midgut of the silkworm, Bombyx mori2005

    • Author(s)
      Sotoshiro, H., Ikeda, M., Tanaka, T., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 74・2

      Pages: 79-82

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Functional analysis of the baculovirus host range gene, hrf-12005

    • Author(s)
      Ikeda, M., Reimbold, E.A., Thiem, S.M.
    • Journal Title

      Virology 332・2

      Pages: 602-613

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Accumulation of non-viral 45.5K and 44K polypeptides inversely correlates with that of viral structural polypeptides in the midgut of the silkworm, Bombyx mori, infected with B. mori densovirus type 2.2005

    • Author(s)
      Sotoshiro, H., Ikeda, M., Tanaka, T., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 74(2)

      Pages: 79-82

    • NAID

      10016610838

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Functional analysis of the baculovirus host range gene, hrf-1.2005

    • Author(s)
      Ikeda, M., Reimbold, E.A., Thiem, S.M.
    • Journal Title

      Virology 332(2)

      Pages: 602-613

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Accumulation of non-viral 45.5K and 44K polypeptides inversely correlates with that of viral structural polypeptides in the midgut of the silkworm, Bombyx mori, infected with B, mori densovirus type 22005

    • Author(s)
      Sotoshiro, H., Ikeda, M., Tanaka, T., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 74・2

      Pages: 79-82

    • NAID

      10016610838

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Identification and functional analysis of Hyphantria cunea nucleopolyhedrovirus iap genes.2004

    • Author(s)
      Ikeda, M., Yanagimoto, K., Kobayashi, M.
    • Journal Title

      Virology 321

      Pages: 359-371

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Association of Sf9 cell proliferating cell nuclear antigen with the DNA replication site of Autographa californica multicapsid nucleopolyhedrovirus.2004

    • Author(s)
      Iwahori, S., Ikeda, M., Kobayashi, M.
    • Journal Title

      Journal of General Virology 85

      Pages: 2857-2862

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Host range factor 1 from Lymantria dispar nucleopolyhedrovirus (NPV) is an essential viral factor required for productive infection of NPVs in IPLB-LD652Y cells derived from L.dispar.2004

    • Author(s)
      Ishikawa, H., Ikeda, M., Felipe Alves, C.A., Thiem, S.M., Kobayashi, M.
    • Journal Title

      Journal of virology 78

      Pages: 12703-12708

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Cloning and characterization of three distinct nucleopolyhedroviruses isolated from the common cutworm, Spodoptera litua in Japan2004

    • Author(s)
      Kamiya, K., Zhu, J., Murata, M., Lavina-Caoili, B.A., et al.
    • Journal Title

      Biological Control 31

      Pages: 38-48

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Restriction of Bombyx mori nucleopolyhedroviruse (NPV) replication in a cell line from B. mori by Hyphantria cunea NPV2004

    • Author(s)
      Shirata, N., Ikeda, M., Kamiya, K., Kawamura, S., Kobayashi, M.
    • Journal Title

      Journal of Insect Biotechnology and Sericology 73

      Pages: 23-33

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Gene organization and complete sequence of the Hyphantria cunea nucleopolyhedrovirus genome

    • Author(s)
      Ikeda, M., Shikata, M., Shirata, N., Chaeychomsri, S., Kobayashi, M.
    • Journal Title

      Journal of General Virology (in press)

    • Related Report
      2005 Annual Research Report
  • [Publications] Ishikawa, H., Ikeda, M., Yanagimoto, K.et al.: "Induction of apoptosis in IPLB-Ld 652Y cell line infected with nucleopolyhedroviruses"J.Geu.Virol.. 84・3. 705-714 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Kamiya, K., Okimoto, N., Ikeda, M.et al.: "Geuotypic variation of a wild isolate of Hyphantria cunea nucleopolyhedrovirus"J.Insect Biotechnol.Sericol.. 72・1. 57-64 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Yamada, T., Ikeda, M., Kobayashi, M., Hattori, K.: "Cloning and expression of a cDNA encoding the laival α-amylase of azuki been weevil Callosobrucus Chinensis"J.Insect Biotechnol.Sericol.. 72・3. 139-148 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ikeda, M., Yanagimoto, K., Kobayashi, M.: "Identification and functional analysis of Hyphantria cunea nucleopholyhedrovirus iap gene"Virology. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Shirata, N., Ikeda, M., Kamiya, K.et al.: "Restriction of Bombyx mori nucleopolyhedrovirus (NPV) replication in a cell line from B.mori by Hyphantria cunea"J.Insect Biotechnol.Sericol.. 73・1(in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Iwahori, S., Ikeda, M, Kobayashi M: "Comparative Characterization of pcnagenes from Hyphantria cunee nucleopolyhedrovirces, and two cell lines from the fall armyworm"J. Insect Biotechnol. Sericol.. 71・1. 25-34 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Iwahori, S., Ikeda, M, Kobayashi M: "Generation and characterization of Autographa californica nucleopolyhedrovirus mutants defective in pcnagene"J. Insect Biotechnol. Sericol.. 71・3. 129-140 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Felipe Alves, C.A., Ikeda, M., Kobayashi, M: "Characterization of Hyphantria cunea NPV gp64 gene and analysis of elements regulating its early promoter activity"J. Insect Biotechnol. Sericol.. 71・3. 141-150 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Felipe Alves, C.A., Ikeda, M., Kobayashi, M: "Identification and characterization of Hyphantria cunea nucleopolyhedrovirus homologues repeated regions"Virus Genes. 25・3. 281-290 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Kamiya, K. 他5名: "Genotypic variation of a wild isolate of Hyphantria cunee nucleopolyhedrovirus"J. Insect Biotechnol. Sericol.. 72・1. 57-64 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ishikawa, H. 他6名: "Induction of apoptosis in an insect cell line, IPLB-Ld652Y, intected with nucleopolyhedroviruses"J. Gen. Virol.. 84・3. 705-714 (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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