The mechanism of genome maintenance and transcriptional regulation in latent HCMV infection

• Project/Area Number: 17590429
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: Aichi Cancer Center Research Institute
• Principal Investigator: ISOMURA Hiroki Department of Virology, Aichi Cancer Center Research Institute, Senior Researcher, 腫瘍ウイルス学部, 主任研究員 (20294415)
• Co-Investigator(Kenkyū-buntansha): 大黒 徹 愛知県がんセンター(研究所), 腫瘍ウイルス学部, 主任研究員 (80291409)
• Project Period (FY): 2005 – 2006
• Project Status: Completed (Fiscal Year 2006)
• Budget Amount: ¥3,600,000 (Direct Cost: ¥3,600,000); Fiscal Year 2006: ¥1,400,000 (Direct Cost: ¥1,400,000); Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: Cytomegalovirus / Transcriptional regulation / 遺伝子制御 / ヒトサイトメガロウイルス / エンハンサー / Sp1 / 前初期遺伝子 / BAC

Research Abstract

We previous demonstrated that the major immediate early (MIE) proximal enhancer containing one GC box and the TATA box containing promoter are minimal elements required for transcription and viral replication (H.Isomura et al. J.Virol. 78:12788, 2004). Here we report that Electrophoretic mobility shift assays (EMSAs) detected binding of Sp1 and Sp3 transcription factors to the GC boxes located at approximately-55 and -75 relative to the transcription start site (+1). Reporter gene experiments showed that both of the Sp1/Sp3 binding sites have a positive and synergistic effect on the HCMV MIE promoter. After infection, the level of Sp1 increased while Sp3 remain constant. There was little to no change in MIE transcription or viral replication for recombinant viruses with one or the other Sp1/Sp3 binding site mutated. In contrast, mutation of both Sp1/Sp3 binding sites caused inefficient MIE transcription and viral replication. These data indicate that the Sp1/3 binding sites have a significant role in HCMV replication.

Research Products

• [Journal Article] Postreplicative mismatch repair factors are recruited to Epstein-Barr virus replication compartments. (2006)
  • Author(s): Daikoku T., Kudoh A., Sugaya Y., Iwahori S., Shirata N., Isomura H., Tsurumi T.
  • Journal Title: J. Biol. Chem. 281 Pages: 11422-11430
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Establishment of a novel foreign gene delivery system combining an HSV amplicon with an attenuated replication-competent virus, HSV-1 HF10. (2006)
  • Author(s): Zhang L., Daikoku T., Ohtake K., Ohtsuka J., Nawa A., Kudoh A., Iwahori S., Isomura H., Nishiyama Y., Tsurumi T
  • Journal Title: J. Virol. Methods 137 Pages: 177-183
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Phosphorylation of MCM4 at sites inactivating DNA helicase activity of the MCM4-MCM6-MCM7 complex during Epstein-Barr virus productive replication. (2006)
  • Author(s): Kudoh A, Daikoku T, Ishimi Y, Kawaguchi Y, Shirata N, Iwahori S, Isomura H, Tsurumi
  • Journal Title: J.Virol. 80
• [Journal Article] Establishment of a novel foreign gene delivery system combining an HSV amplicon with an attenuated replication-competent virus, HSV-1 HSV-1 HF10. (2006)
  • Author(s): Zhang L, Daikoku T, Ohatake K, Ohtsuka J, Nawa A, Kudoh A, Iwahori S, Isomura H, Nishiyama Y, Tsurumi T.
  • Journal Title: J.Virol.Methods 137 Pages: 177-183
• [Journal Article] Postreplicative mismatch repair factors are recruited to Epstein-Barr virus replication compartments. (2006)
  • Author(s): Daikoku T, Kudoh A, Sugaya Y, Iwahori S, Shirata N, Isomura H, Tsurumi T.
  • Journal Title: J.Biol.Chem. 281 Pages: 11422-30
• [Journal Article] Activation of ataxia telangiectasia-mutated DNA damage checkpoint signal transduction elicited by herpes simplex virus infection. (2006)
  • Author(s): Shirata N, Kudoh A, Daikoku T, Tatsumi Y, Fujita M, Kiyono T, Sugaya Y, Isomura H, Ishizaki K, Tsurumi T.
  • Journal Title: J.Biol.Chem. 280 Pages: 30336-30341
• [Journal Article] Two Sp1/Sp3 Binding Sites in the Proximal Enhancer of the Human Cytomegalovirus Major Immediate-Early Genes are necessary for its transcriptional activation. (2005)
  • Author(s): Isomura H., Stinski M.F., Kudoh A., Daikoku T., Shirata N., Tsurumi T.
  • Journal Title: J. Virol. 79 Pages: 9597-9607
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Epstein-Barr virus lytic replication elicits ATM checkpoint signal transduction while providing an S-phase-like cellular environment. J. Biol. Chem., 280 : 8156-8163, 2005 (2005)
  • Author(s): Kudoh A., Fujita M., Zhan L., Shirata N., Daikoku T., Sugaya Y., Isomura H., Nishiyama Y., Tsurumi T
  • Journal Title: J. Biol. Chem. 280 Pages: 8156-8163
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Architecture of replication compartments formed during Epstein-Barr virus lytic replication. (2005)
  • Author(s): Daikoku T., Kudoh A., Fujita M., Sugaya Y., Isomura H., Shirata N., Tsurumi T
  • Journal Title: J. Virol. 79 Pages: 3409-3418
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Two Sp1/Sp3 Binding Sites in the Proximal Enhancer of the Human Cytomegalovirus Major Immediate-Early Genes are necessary for its transcriptional activation. (2005)
  • Author(s): Isomura H., Stinski M.F., Kudoh A., Daikoku T., Shirata N., Tsurumi T.
  • Journal Title: J.Virol. 79 Pages: 9597-9607
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Epstein-Barr virus lytic replication elicits ATM checkpoint signal transduction while providing an S-phase-like cellular environment. (2005)
  • Author(s): Kudoh A., Fujita M., Zhan L., Shirata N., Daikoku T., Sugaya Y., Isomura H., Nishiyama Y., Tsurumi T.
  • Journal Title: J.Biol.Chem. 280 Pages: 8156-8163
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Architecture of replication compartments formed during Epstein-Barr virus lytic replication. (2005)
  • Author(s): Daikoku T., Kudoh A., Fujita M., Sugaya Y., Isomura H., Shirata N., Tsurumi T.
  • Journal Title: J.Virol. 79 Pages: 3409-3418
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Two Sp1/Sp3 binding sites in the major immediate-early proximal enhancer of human cytomegalovirus have a significant role in viral replication. (2005)
  • Author(s): Isomura, H., Stinski, M.F., Kudoh, A., Daikoku, T., Shirata, N., Tsurumi
  • Journal Title: J.Virol. 79 Pages: 9597-9607
• [Journal Article] Architecture of replication compartments formed during Epstein-Barr virus lytic replication (2005)
  • Author(s): Daikoku, T., Kudoh, A., Fujita, M., Sugaya, Y., Isomura, H., Shirata, N., Tsurumi, T
  • Journal Title: J.Virol. 79 Pages: 3409-3418
• [Journal Article] Activation of ataxia telangiectasia-mutated DNA damage checkpoint signal transduction elicited by herpes simplex virus infection (2005)
  • Author(s): Shirata, N., Kudoh, A., Daikoku, T., Tatsumi, Y., Fujita, M., Kiyono, T., Sugaya, Y., Isomura, H., Ishizaki, K., Tsurumi, T
  • Journal Title: J.Biol.Chem 280 Pages: 30336-30341
• [Book] ヘルペスウイルスの組織親和性 (2006)
  • Author(s): 磯村寛樹, 鶴見達也
  • Publisher: 日本臨床社
  • Description: 「研究成果報告書概要(和文)」より
• [Book] ヘルペスウイルスの組織親和性 (2006)
  • Author(s): 磯村寛樹, 鶴見達也
  • Publisher: 日本臨床社,日本臨床増刊号
• [Patent(Industrial Property Rights)] 弱毒化組換えヒトサイトメガロウイルス (2006)
  • Inventor(s): 磯村 寛樹, 鶴見 達也
  • Industrial Property Number: 2006-277793
  • Filing Date: 2006-10-11
• [Patent(Industrial Property Rights)] 弱毒化組換えヒトサイトメガロウイルス
  • Inventor(s): 磯村寛樹
  • Industrial Property Rights Holder: 特願2006-277793
  • Description: 「研究成果報告書概要(和文)」より