U.S. Department of Energy

Pacific Northwest National Laboratory

Release of severe acute respiratory syndrome coronavirus nuclear import block enhances host transcription in human lung cells.

TitleRelease of severe acute respiratory syndrome coronavirus nuclear import block enhances host transcription in human lung cells.
Publication TypeJournal Article
Year of Publication2013
AuthorsSims AC, Tilton SC, Menachery VD, Gralinski LE, Schäfer A, Matzke MM, Webb-Robertson B-JM, Chang J, Luna ML, Long CE, Shukla AK, Bankhead AR, Burkett SE, Zornetzer G, Tseng C-TKent, Metz TO, Pickles R, McWeeney S, Smith RD, Katze MG, Waters KM, Baric RS
JournalJ Virol
KeywordsActive Transport, Cell Nucleus, Chromatin Immunoprecipitation, Computational Biology, DNA Primers, Epithelial Cells, Gene Regulatory Networks, Humans, Lung, Microarray Analysis, Proteomics, Real-Time Polymerase Chain Reaction, SARS Virus, Signal Transduction, Systems Biology, Transcription, Genetic, Viral Regulatory and Accessory Proteins
Abstract

The severe acute respiratory syndrome coronavirus accessory protein ORF6 antagonizes interferon signaling by blocking karyopherin-mediated nuclear import processes. Viral nuclear import antagonists, expressed by several highly pathogenic RNA viruses, likely mediate pleiotropic effects on host gene expression, presumably interfering with transcription factors, cytokines, hormones, and/or signaling cascades that occur in response to infection. By bioinformatic and systems biology approaches, we evaluated the impact of nuclear import antagonism on host expression networks by using human lung epithelial cells infected with either wild-type virus or a mutant that does not express ORF6 protein. Microarray analysis revealed significant changes in differential gene expression, with approximately twice as many upregulated genes in the mutant virus samples by 48 h postinfection, despite identical viral titers. Our data demonstrated that ORF6 protein expression attenuates the activity of numerous karyopherin-dependent host transcription factors (VDR, CREB1, SMAD4, p53, EpasI, and Oct3/4) that are critical for establishing antiviral responses and regulating key host responses during virus infection. Results were confirmed by proteomic and chromatin immunoprecipitation assay analyses and in parallel microarray studies using infected primary human airway epithelial cell cultures. The data strongly support the hypothesis that viral antagonists of nuclear import actively manipulate host responses in specific hierarchical patterns, contributing to the viral pathogenic potential in vivo. Importantly, these studies and modeling approaches not only provide templates for evaluating virus antagonism of nuclear import processes but also can reveal candidate cellular genes and pathways that may significantly influence disease outcomes following severe acute respiratory syndrome coronavirus infection in vivo.

DOI10.1128/JVI.02520-12
Alternate JournalJ. Virol.
PubMed ID23365422
PubMed Central IDPMC3624188
Grant ListHHSN272200800060C / AO / NIAID NIH HHS / United States
UL1 TR000128 / TR / NCATS NIH HHS / United States
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