heparitin-sulfate and Vaccinia

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/p24δ) 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.

Topics: Cell Line, Tumor; Cowpox virus; CRISPR-Cas Systems; DNA Viruses; Gene Knockout Techniques; Genetic Testing; Golgi Apparatus; Haploidy; HEK293 Cells; HeLa Cells; Heparitin Sulfate; Host Specificity; Host-Pathogen Interactions; Humans; Membrane Proteins; Monkeypox virus; N-Acetylglucosaminyltransferases; Phosphatidylserines; Pinocytosis; Poxviridae; Vaccinia; Vaccinia virus; Vesicular Transport Proteins; Virus Attachment