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

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

  • Mol Cell. 2020 May 21;78(4):624-640.e7. doi: 10.1016/j.molcel.2020.04.013.
Byungil Kim 1,  Sarah Arcos 1,  Katherine Rothamel 1,  Jeffrey Jian 1,  Kristie L Rose 2,  W Hayes McDonald 2,  Yuqi Bian 1,  Seth Reasoner 1,  Nicholas J Barrows 3,  Shelton Bradrick 3,  Mariano A Garcia-Blanco 4,  Manuel Ascano 5
Affiliations

Affiliations

  • 1 Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
  • 2 Department of Biochemistry and Mass Spectrometry Research Center, Vanderbilt University, Nashville, TN 37232, USA.
  • 3 Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, TX 77555, USA.
  • 4 Department of Biochemistry and Molecular Biology, The University of Texas Medical Branch, Galveston, TX 77555, USA; Programme in Emerging Infectious Diseases, Duke-NUS Medical School, Singapore 169857, Singapore.
  • 5 Department of Biochemistry, Vanderbilt University School of Medicine, Nashville, TN 37232, USA. Electronic address: manuel.ascano@vanderbilt.edu.
PMID: 32380061 DOI: 10.1016/j.molcel.2020.04.013
Abstract

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 explore 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 fatty acid synthase (FASN) for pro-viral activity. We show that CHIKV genomes are N6-methyladenosine modified, and YTHDF1 binds and suppresses CHIKV 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.

Keywords

RNA virus; RNA-binding protein; VIR-CLASP; host-pathogen interactions; innate immunity; interactome capture.

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