Developing an anti-HIV drugs that inhibit uncoating for overcomingdrug resistance

2012 Fiscal Year Final Research Report

• Project/Area Number: 23659066
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Environmental pharmacy
• Research Institution: Kumamoto University
• Principal Investigator: MISUMI Shogo 熊本大学, 大学院・生命科学研究部, 准教授 (40264311)
• Project Period (FY): 2011 – 2012
• Keywords: ウイルス / 感染症 / 微生物

Research Abstract

We previously reported that Pin1 facilitates the humanimmunodeficiency virus type 1 (HIV -1) uncoating by interacting with the capsid (CA) corethrough the phosphorylated Ser16-Pro17motif. However , the specific kinase responsible forSer16phosphorylation has remained unknown. Here, we show that virion-associatedextracellular signal-regulated kinase 2 (ERK2) phosphorylates Ser16. The characterization ofimmature virions produced by exposing of CEM/LA V -1 cells to 10 μM saquinavir and a D25Ninactive mutant of HIV -1 protease indicated that Ser16is phosphorylated after the initiationof Pr55gagprocessing. Furthermore, a mass spectrometry-based in vitro kinase assaydemonstrated that ERK2 specifically phosphorylated the Ser16residue in theSer16-Pro17-motif-containing substrate. The treatment of CEM/LA V -1 cells with the ERK2inhibitor sc-222229 decreased the Ser16phosphorylation level inside virions, and the viruspartially defective in Ser16phosphorylation showed an impaired reverse transcription and anattenuated replication owing to an attenuated Pin1-dependent uncoating. Furthermore, thesuppression of ERK2 expression by RNA interference in CEM/LA V -1 cells resulted insuppressed ERK2 packaging inside virions and decreased the Ser16phosphorylation levelinside virions. Interestingly , the ERK2-packaging-defective virus showed an impaired reversetranscription and an attenuated HIV -1 replication. Taken together , these findings provideinsights into the as yet obscure processes in Pin1-dependent HIV -1 uncoating.

Research Products

• [Journal Article] Induction of extremely low proteinexpression level by fusion ofC-terminal region of Nef (2012)
  • Author(s): Takamune, N.*, Irisaka, Y., Yamamoto,M., Harada, K., Shoji, S., Misumi, S
  • Journal Title: Biotechnology and AppliedBiochemistry Volume: 59 Pages: 245-253
  • Peer Reviewed
• [Journal Article] Glyceraldehyde 3-phosphatedehydrogenase negatively regulateshuman immunodeficiency virus type 1infection (2012)
  • Author(s): Kishimoto, N., Onitsuka, A., Kido, K.,Takamune, N., Shoji, S., Misumi, S
  • Journal Title: Retrovirology Volume: 9 Pages: 107
  • Peer Reviewed
• [Presentation] HIV 脱殻過程を制御する HIV カプシドタンパク質リン酸化機構の解明 (2012)
  • Author(s): 堂地 赳生、高宗 暢暁、庄司 省三、杉本 幸彦、三隅 将吾
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: 福岡国際会議場(福岡)
  • Year and Date: 20121214-1216
• [Presentation] ERK2 による HIV CA の Ser16 リン酸化を介した脱殻制御機構に関する解析 (2012)
  • Author(s): 堂地 赳生、高宗 暢暁、庄司 省三、杉本 幸彦、三隅 将吾
  • Organizer: 第26回 日本エイズ学会学術集会・総会
  • Place of Presentation: 慶應義塾大学 日吉キャンパス(横浜)
  • Year and Date: 20121124-1126
• [Presentation] HIV-1 脱殻素過程を制御するカプシド蛋白質 Ser16 リン酸化機構に関する解析 (2012)
  • Author(s): 堂地 赳生、高宗 暢暁、庄司 省三、杉本 幸彦、三隅 将吾
  • Organizer: 第60回 日本ウイルス学会学術集会プログラム
  • Place of Presentation: 大阪国際会議場(大阪)
  • Year and Date: 20121113-1115
• [Presentation] HIV-1 カプシド蛋白質 Ser16 リン酸化による脱殻素過程の制御機構堂地 (2012)
  • Author(s): 堂地 赳生、井上 睦美、高宗 暢暁、庄司 省三、杉本 幸彦、三隅 将吾
  • Organizer: 次世代を担う若手ファーマ ・バイオフォーラム
  • Place of Presentation: 九州大学病院キャンパス(福岡)
  • Year and Date: 20120915-0916