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DOI	10.1073/pnas.2010524118
	Measuring the subcellular compartmentalization of viral infections by protein complementation assay
	Fernandez J.; Hassen-Khodja C.; Georget V.; Rose T.; Jacob Y.; Janin Y.L.; Nisole S.; Vidalain P.-O.; Arhel N.J.
发表日期	2021
ISSN	00278424
卷号	118 期号:2
英文摘要	The recent emergence and reemergence of viruses in the human population has highlighted the need to develop broader panels of therapeutic molecules. High-throughput screening assays opening access to untargeted steps of the viral replication cycle will provide powerful leverage to identify innovative antiviral molecules. We report here the development of an innovative protein complementation assay, termed αCentauri, to measure viral translocation between subcellular compartments. As a proof of concept, the Centauri fragment was either tethered to the nuclear pore complex or sequestered in the nucleus, while the complementary α fragment (<16 amino acids) was attached to the integrase proteins of infectious HIV-1. The translocation of viral ribonucleoproteins from the cytoplasm to the nuclear envelope or to the nucleoplasm efficiently reconstituted superfolder green fluorescent protein or NanoLuc αCentauri reporters. These fluorescence- or bioluminescence-based assays offer a robust readout of specific steps of viral infection in a multiwell format that is compatible for high-throughput screening and is validated by a short hairpin RNA-based prototype screen. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Subcellular compartments; Technological development; Viruses
语种	英语
scopus关键词	alpha centauri protein; enhanced green fluorescent protein; integrase; karyopherin beta; ribonucleoprotein; short hairpin RNA; unclassified drug; viral protein; Article; carboxy terminal sequence; cell compartmentalization; cell nucleus membrane; controlled study; cytoplasm; Human immunodeficiency virus 1; Human immunodeficiency virus 1 infection; hydrolysis; long terminal repeat; nonhuman; nuclear import; nuclear localization signal; priority journal; protein expression; protein localization; protein stability; protein-fragment complementation assay; signal noise ratio; virus capsid; virus entry; virus pathogenesis; virus replication
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/181070
作者单位	Institut de Recherche en Infectiologie de Montpellier, CNRS, UMR 9004, Université de Montpellier, Montpellier, 34090, France; Montpellier Ressources Imagerie, BioCampus Montpellier, CNRS, INSERM, Université de Montpellier, Montpellier, 34090, France; Unité de Biologie Cellulaire des Lymphocytes, Institut Pasteur, INSERM 1221, Paris, 75015, France; Unité de Génétique Moléculaire des Virus à Acide Ribonucléique, Institut Pasteur, CNRS, UMR 3569, Paris, 75015, France; Unité de Chimie et Biocatalyse, Institut Pasteur, CNRS, UMR 3523, Paris, 75724, France; Centre International de Recherche en Infectiologie, Université de Lyon, INSERM U1111, Université Claude Bernard Lyon 1, CNRS, UMR 5308, École Normale Supérieure de Lyon, Lyon, 69342, France; Equipe Chimie et Biologie, Modélisation et Immunologie pour la Thérapie, Université Paris Descartes, CNRS, UMR 8601, Paris, 75006, France
推荐引用方式 GB/T 7714	Fernandez J.,Hassen-Khodja C.,Georget V.,et al. Measuring the subcellular compartmentalization of viral infections by protein complementation assay[J],2021,118(2).
APA	Fernandez J..,Hassen-Khodja C..,Georget V..,Rose T..,Jacob Y..,...&Arhel N.J..(2021).Measuring the subcellular compartmentalization of viral infections by protein complementation assay.Proceedings of the National Academy of Sciences of the United States of America,118(2).
MLA	Fernandez J.,et al."Measuring the subcellular compartmentalization of viral infections by protein complementation assay".Proceedings of the National Academy of Sciences of the United States of America 118.2(2021).