Please use this identifier to cite or link to this item: https://doi.org/10.1186/s12977-018-0407-4
Title: An RNA-binding compound that stabilizes the HIV-1 gRNA packaging signal structure and specifically blocks HIV-1 RNA encapsidation
Authors: Ingemarsdotter, C.K
Zeng, J
Long, Z
Lever, A.M.L 
Kenyon, J.C 
Keywords: 4 [(1 methyl 6 nitro 4(1h) quinolinylidene)amino] n [4 [(1 methyl 4(1h) pyridinylidene)amino]phenyl]benzamide
antiretrovirus agent
Gag protein
genomic RNA
nsc 260594
unclassified drug
virus RNA
protein binding
5' untranslated region
Article
conformational transition
controlled study
cytoplasm
dimerization
enzyme linked immunosorbent assay
Human immunodeficiency virus 1
nonhuman
reverse transcription polymerase chain reaction
RNA encapsidation
RNA structure
virion
virus DNA cell DNA interaction
virus entry
virus infectivity
virus particle
Western blotting
chemistry
conformation
genetics
genomic instability
human
Human immunodeficiency virus 1
Human immunodeficiency virus infection
metabolism
physiology
provirus
real time polymerase chain reaction
virology
virus assembly
virus genome
virus load
virus release
5' Untranslated Regions
Gene Products, gag
Genome, Viral
Genomic Instability
HIV Infections
HIV-1
Humans
Nucleic Acid Conformation
Protein Binding
Proviruses
Real-Time Polymerase Chain Reaction
RNA, Viral
Viral Load
Virus Assembly
Virus Integration
Virus Release
Issue Date: 2018
Citation: Ingemarsdotter, C.K, Zeng, J, Long, Z, Lever, A.M.L, Kenyon, J.C (2018). An RNA-binding compound that stabilizes the HIV-1 gRNA packaging signal structure and specifically blocks HIV-1 RNA encapsidation. Retrovirology 15 (1) : 25. ScholarBank@NUS Repository. https://doi.org/10.1186/s12977-018-0407-4
Rights: Attribution 4.0 International
Abstract: Background: NSC260594, a quinolinium derivative from the NCI diversity set II compound library, was previously identified in a target-based assay as an inhibitor of the interaction between the HIV-1 (?) stem-loop 3 (SL3) RNA and Gag. This compound was shown to exhibit potent antiviral activity. Here, the effects of this compound on individual stages of the viral lifecycle were examined by qRT-PCR, ELISA and Western blot, to see if its actions were specific to the viral packaging stage. The structural effects of NSC260594 binding to the HIV-1 gRNA were also examined by SHAPE and dimerization assays. Results: Treatment of cells with NSC260594 did not reduce the number of integration events of incoming virus, and treatment of virus producing cells did not affect the level of intracellular Gag protein or viral particle release as determined by immunoblot. However, NSC260594 reduced the incorporation of gRNA into virions by up to 82%, without affecting levels of gRNA inside the cell. This reduction in packaging correlated closely with the reduction in infectivity of the released viral particles. To establish the structural effects of NSC260594 on the HIV-1 gRNA, we performed SHAPE analyses to pinpoint RNA structural changes. NSC260594 had a stabilizing effect on the wild type RNA that was not confined to SL3, but that was propagated across the structure. A packaging mutant lacking SL3 did not show this effect. Conclusions: NSC260594 acts as a specific inhibitor of HIV-1 RNA packaging. No other viral functions are affected. Its action involves preventing the interaction of Gag with SL3 by stabilizing this small RNA stem-loop which then leads to stabilization of the global packaging signal region (psi or ?). This confirms data, previously only shown in analyses of isolated SL3 oligonucleotides, that SL3 is structurally labile in the presence of Gag and that this is critical for the complete psi region to be able to adopt different conformations. Since replication is otherwise unaffected by NSC260594 the flexibility of SL3 appears to be a unique requirement for genome encapsidation and identifies this process as a highly specific drug target. This study is proof of principle that development of a new class of antiretroviral drugs that specifically target viral packaging by binding to the viral genomic RNA is achievable. © 2018 The Author(s).
Source Title: Retrovirology
URI: https://scholarbank.nus.edu.sg/handle/10635/178100
ISSN: 17424690
DOI: 10.1186/s12977-018-0407-4
Rights: Attribution 4.0 International
Appears in Collections:Staff Publications
Elements

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
10_1186_s12977-018-0407-4.pdf1.78 MBAdobe PDF

OPEN

NoneView/Download

Google ScholarTM

Check

Altmetric


This item is licensed under a Creative Commons License Creative Commons