A Genome-Wide Haploid Genetic Screen Identifies Heparan Sulfate-Associated Genes and the Macropinocytosis Modulator TMED10 as Factors Supporting Vaccinia Virus Infection
| Autor: | Toin H. van Kuppevelt, Emmanuel J. H. J. Wiertz, Rutger D. Luteijn, Ingo Drexler, Robert Jan Lebbink, Ingrid G. J. Boer, Vincent A. Blomen, Ferdy R. van Diemen, Saravanan Manikam Sadasivam, Thijn R. Brummelkamp |
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| Rok vydání: | 2019 |
| Předmět: |
viruses
Vesicular Transport Proteins Golgi Apparatus Heparan sulfate Haploidy TMED10 chemistry.chemical_compound Gene Knockout Techniques Vaccinia Monkeypox virus Phosphatidylserine biology Cowpox virus Virus-Cell Interactions Reconstructive and regenerative medicine Radboud Institute for Molecular Life Sciences [Radboudumc 10] Poxvirus Host-Pathogen Interactions Variola virus Immunology Virus Attachment Vaccinia virus Phosphatidylserines N-Acetylglucosaminyltransferases Microbiology Virus Host Specificity Insertional mutagenesis Cell Line Tumor Virology Humans Genetic Testing Tropism Macropinocytosis Genome-wide screen Poxviridae DNA Viruses Membrane Proteins biology.organism_classification HEK293 Cells chemistry Insect Science Pinocytosis Heparitin Sulfate CRISPR-Cas Systems HeLa Cells |
| Zdroj: | Journal of Virology Journal of Virology, 93(13). American Society for Microbiology Journal of Virology, 93 Journal of Virology, 93, 13 |
| ISSN: | 0022-538X |
| Popis: | Contains fulltext : 205154.pdf (Publisher’s version ) (Closed access) Vaccinia virus is a promising viral vaccine and gene delivery candidate and has historically been used as a model to study poxvirus-host cell interactions. We employed a genome-wide insertional mutagenesis approach in human haploid cells to identify host factors crucial for vaccinia virus infection. A library of mutagenized HAP1 cells was exposed to modified vaccinia virus Ankara (MVA). Deep-sequencing analysis of virus-resistant cells identified host factors involved in heparan sulfate synthesis, Golgi organization, and vesicular protein trafficking. We validated EXT1, TM9SF2, and TMED10 (TMP21/p23/p24delta) as important host factors for vaccinia virus infection. The critical roles of EXT1 in heparan sulfate synthesis and vaccinia virus infection were confirmed. TM9SF2 was validated as a player mediating heparan sulfate expression, explaining its contribution to vaccinia virus infection. In addition, TMED10 was found to be crucial for virus-induced plasma membrane blebbing and phosphatidylserine-induced macropinocytosis, presumably by regulating the cell surface expression of the TAM receptor Axl.IMPORTANCE Poxviruses are large DNA viruses that can infect a wide range of host species. A number of these viruses are clinically important to humans, including variola virus (smallpox) and vaccinia virus. Since the eradication of smallpox, zoonotic infections with monkeypox virus and cowpox virus are emerging. Additionally, poxviruses can be engineered to specifically target cancer cells and are used as a vaccine vector against tuberculosis, influenza, and coronaviruses. Poxviruses rely on host factors for most stages of their life cycle, including attachment to the cell and entry. These host factors are crucial for virus infectivity and host cell tropism. We used a genome-wide knockout library of host cells to identify host factors necessary for vaccinia virus infection. We confirm a dominant role for heparin sulfate in mediating virus attachment. Additionally, we show that TMED10, previously not implicated in virus infections, facilitates virus uptake by modulating the cellular response to phosphatidylserine. |
| Databáze: | OpenAIRE |
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