Proteomics

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Discovery of Widespread Host Protein Interactions with the Pre-Replicated Genome of CHIKV using VIR-CLASP


ABSTRACT: The pioneer interactions between incoming viral RNA genomes and host proteins are crucial to infection and immune response. Until now, the ability to study these events was lacking. We developed VIR-CLASP (VIRal Cross-Linking And Solid-phase Purification) to characterize the earliest interactions between viral RNA and cellular proteins. We investigated the infection of human cells using Chikungunya virus (CHIKV) and Influenza A virus and identified hundreds of direct RNA-protein interactions. Here, we validate the biological impact of three protein classes that bind CHIKV RNA within minutes of infection. We find CHIKV RNA binds and hijacks the lipid-modifying enzyme FASN for pro-viral activity. We show that CHIKV genomes are N6-methyladenosine modified and that YTHDF1 binds and suppresses its replication. Finally, we find that the innate immune DNA sensor IFI16 associates with CHIKV RNA, reducing viral replication and maturation. Our findings have direct applicability to the investigation of potentially all RNA viruses.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

SUBMITTER: Manny Ascano  

LAB HEAD: Manuel Ascano

PROVIDER: PXD015863 | Pride | 2020-05-11

REPOSITORIES: Pride

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Publications

Discovery of Widespread Host Protein Interactions with the Pre-replicated Genome of CHIKV Using VIR-CLASP.

Kim Byungil B   Arcos Sarah S   Rothamel Katherine K   Jian Jeffrey J   Rose Kristie L KL   McDonald W Hayes WH   Bian Yuqi Y   Reasoner Seth S   Barrows Nicholas J NJ   Bradrick Shelton S   Garcia-Blanco Mariano A MA   Ascano Manuel M  

Molecular cell 20200506 4


The primary interactions between incoming viral RNA genomes and host proteins are crucial to infection and immunity. Until now, the ability to study these events was lacking. We developed viral cross-linking and solid-phase purification (VIR-CLASP) to characterize the earliest interactions between viral RNA and cellular proteins. We investigated the infection of human cells using Chikungunya virus (CHIKV) and influenza A virus and identified hundreds of direct RNA-protein interactions. Here, we  ...[more]

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