ArticleJournal of virology2013
Thiouracil cross-linking mass spectrometry: a cell-based method to identify host factors involved in viral amplification.
Article in Journal of virology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 36 papers.
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Who cites it
36 citing papers in PubMed.
- Subversion of phosphorylated SR proteins by enterovirus A71 in IRES-dependent translation revealed by RNA-interactome analysis.PLoS pathogens · 2025Article
- Cellular NONO protein binds to the flavivirus replication complex and promotes positive-strand RNA synthesis.Journal of virology · 2024Article
- Involvement of paraspeckle components in viral infections.Nucleus (Austin, Tex.) · 2024Review
- Review
- Viral RNA Is a Hub for Critical Host-Virus Interactions.Sub-cellular biochemistry · 2023Article
- A Proximity biotinylation assay with a host protein bait reveals multiple factors modulating enterovirus replication.PLoS pathogens · 2022Article
- Uncovering viral RNA-host cell interactions on a proteome-wide scale.Trends in biochemical sciences · 2022Review
- Pathogenesis and virulence of flavivirus infections.Virulence · 2021Review
- RNA Binding Proteins as Pioneer Determinants of Infection: Protective, Proviral, or Both?Viruses · 2021Review
- Article
- Article
- The importance of virion-incorporated cellular RNA-Binding Proteins in viral particle assembly and infectivity.Seminars in cell & developmental biology · 2021Review
- RNA-Centric Approaches to Profile the RNA-Protein Interaction Landscape on Selected RNAs.Non-coding RNA · 2021Review
- Systematic discovery and functional interrogation of SARS-CoV-2 viral RNA-host protein interactions during infection.bioRxiv : the preprint server for biology · 2020Article
- Discovery of Widespread Host Protein Interactions with the Pre-replicated Genome of CHIKV Using VIR-CLASP.Molecular cell · 2020Article
- Unconventional RNA-binding proteins step into the virus-host battlefront.Wiley interdisciplinary reviews. RNA · 2018Review
- Article
- Exploitation of nuclear functions by human rhinovirus, a cytoplasmic RNA virus.PLoS pathogens · 2018Article
- Identification and Characterization of Sindbis Virus RNA-Host Protein Interactions.Journal of virology · 2018Article
- Aedes aegypti microRNA miR-2b regulates ubiquitin-related modifier to control chikungunya virus replication.Scientific reports · 2017Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Eukaryotic RNA viruses are known to utilize host factors; however, the identity of these factors and their role in the virus life cycle remain largely undefined. Here, we report a method to identify proteins bound to the viral RNA during amplification in cell culture: thiouracil cross-linking mass spectrometry (TUX-MS). TUX-MS relies on incorporation of a zero-distance cross-linker into the viral RNA during infection. Proteins bound to viral RNA are cross-linked prior to cell lysis, purified, and identified using mass spectrometry. Using the TUX-MS method, an unbiased screen for poliovirus (PV) host factors was conducted. All host and viral proteins that are known to interact with the poliovirus RNA were identified. In addition, TUX-MS identified an additional 66 host proteins that have not been previously described in poliovirus amplification. From these candidates, eight were selected and validated. Furthermore, we demonstrate that small interfering RNA (siRNA)-mediated knockdown of two of these uncharacterized host factors results in either a decrease in copy number of positive-stranded RNA or a decrease in PV translation. These data demonstrate that TUX-MS is a robust, unbiased method to identify previously unknown host cell factors that influence virus growth. This method is broadly applicable to a range of RNA viruses, such as flaviviruses, alphaviruses, picornaviruses, bunyaviruses, and coronaviruses.
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