title: [Establishment and application of a high-throughput screening assay for premature activation of HIV-1 precursors].
abstract: Strict regulation of HIV-1 PR function is critical for efficient production of mature viral particles. During viral protein expression and viral assembly, HIV-1 PR located within Gag-Pol precursor must be inactive to prevent premature cytoplasmic processing of the viral Gag and Gag-Pol precursors. Premature activation of HIV-1 precursors leads to major defects in viral assembly and production of viral particles. A cell-level premature activation of HIV-1 precursors assay using bioluminescence resonance energy transfer (BRET) was established. Three thousand compounds were screened to evaluate this assay. The results showed that the assay is sensitive, specific and stable (Z' factor is 0.905).
authors.affiliation: Department of Immunology, Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100050, China.
authors.forename: Quan
authors.initials: Q
authors.lastname: Zhang
authors.forename: Xiao-yu
authors.initials: XY
authors.lastname: Li
authors.forename: Zhen-long
authors.initials: ZL
authors.lastname: Liu
authors.forename: Ping-ping
authors.initials: PP
authors.lastname: Jia
authors.forename: Xiao-lu
authors.initials: XL
authors.lastname: Wei
authors.forename: Li-xun
authors.initials: LX
authors.lastname: Zhao
authors.forename: Jian-dong
authors.initials: JD
authors.lastname: Jiang
authors.forename: Shan
authors.initials: S
authors.lastname: Cen
mesh_terms: D000480:Alkynes; D019380:Anti-HIV Agents / Q000494:pharmacology*; D048588:Benzoxazines / Q000494:pharmacology; D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods; D003521:Cyclopropanes; D015685:Fusion Proteins, gag-pol / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D016333:HIV Protease / Q000378:metabolism* / Q000502:physiology; D015497:HIV-1 / Q000201:enzymology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D009570:Nitriles; D010957:Plasmids / Q000235:genetics; D011498:Protein Precursors / Q000378:metabolism* / Q000502:physiology; D011724:Pyridazines / Q000494:pharmacology; D011743:Pyrimidines; D014162:Transfection; D014771:Virion / Q000254:growth & development; D019065:Virus Assembly; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000235:genetics / Q000378:metabolism
chemical_list: D000480:Alkynes; D019380:Anti-HIV Agents; D048588:Benzoxazines; D003521:Cyclopropanes; D015685:Fusion Proteins, gag-pol; D009570:Nitriles; D011498:Protein Precursors; D011724:Pyridazines; D011743:Pyrimidines; D054301:gag Gene Products, Human Immunodeficiency Virus; C451734:etravirine; D016333:HIV Protease; C098320:efavirenz