@article{b54d961458af40d9b42b99bef2e2d158,
title = "Discovery of Widespread Host Protein Interactions with the Pre-replicated Genome of CHIKV Using VIR-CLASP",
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",
author = "Byungil Kim and Sarah Arcos and Katherine Rothamel and Jeffrey Jian and Rose, {Kristie L.} and McDonald, {W. Hayes} and Yuqi Bian and Seth Reasoner and Barrows, {Nicholas J.} and Shelton Bradrick and Garcia-Blanco, {Mariano A.} and Manuel Ascano",
note = "Funding Information: We would like to thank Matthew Albertolle for technical help in the MS analysis; Neil Sprenkle and James Held for technical help in the molecular studies; Dr. Thomas Voss and Dr. James E. Crowe Jr. (Vanderbilt University Medical Center) for IAV, VSV, and RVFV; Dr. Mark R. Denison (Vanderbilt University Medical Center) for MHV; and Dr. Terence S. Dermody (University of Pittsburgh School of Medicine) for CHIKV. Finally, we would like to thank members of the Ascano laboratory for their support, collegiality, and critical review of the manuscript. This work was supported by the NIH (R35GM119569 to M.A.), the National Center for Advancing Translational Sciences (CTSA) (UL1TR000445 to B.K. and M.A.), Vanderbilt University Department of Biochemistry start-up funds (M.A.), the Chemical Biology of Infectious Disease (training grant 5T32AI11254-02 to S.A.), and the Chemistry-Biology Interface (training grant 5T32GM065086-14). Conceptualization & Methodology, B.K. and M.A.; Software and Data Curation, S.A.; Formal Analysis, S.A. K.R. K.L.R. and W.H.M.; Investigation, B.K. S.A. Y.B. J.J. K.R. and S.R.; Resources, S.B. N.J.B. and M.A.G.-B.; Writing – Original Draft, S.A.; Writing – Review & Editing, B.K. M.A. K.L.R. and W.H.M.; Visualization, S.A. and B.K.; Funding Acquisition, M.A.; Supervision, M.A. M.A.G.-B. K.L.R. and W.H.M. The authors declare no competing interests. Funding Information: We would like to thank Matthew Albertolle for technical help in the MS analysis; Neil Sprenkle and James Held for technical help in the molecular studies; Dr. Thomas Voss and Dr. James E. Crowe Jr. (Vanderbilt University Medical Center) for IAV, VSV, and RVFV; Dr. Mark R. Denison (Vanderbilt University Medical Center) for MHV; and Dr. Terence S. Dermody (University of Pittsburgh School of Medicine) for CHIKV. Finally, we would like to thank members of the Ascano laboratory for their support, collegiality, and critical review of the manuscript. This work was supported by the NIH ( R35GM119569 to M.A.), the National Center for Advancing Translational Sciences (CTSA) ( UL1TR000445 to B.K. and M.A.), Vanderbilt University Department of Biochemistry start-up funds (M.A.), the Chemical Biology of Infectious Disease (training grant 5T32AI11254-02 to S.A.), and the Chemistry-Biology Interface (training grant 5T32GM065086-14 ). Publisher Copyright: {\textcopyright} 2020 Elsevier Inc.",
year = "2020",
month = may,
day = "21",
doi = "10.1016/j.molcel.2020.04.013",
language = "English (US)",
volume = "78",
pages = "624--640.e7",
journal = "Molecular cell",
issn = "1097-2765",
publisher = "Cell Press",
number = "4",
}