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Custom Vector Construction> | Codon-optimized MERS S-containing sequences for a C-terminal C9 epitope tag were purchased from Genscript and subsequently cloned into pcDNA3.1+ between the EcoRI and NotI restriction sites.CD9 and scramble control short hairpin RNA (shRNA) constructs flanked by the U6 promoter and a RNA polymerase III stop sequence were engineered into the pUC57 vector by Genscript. | Get A Quote |
Coronaviruses (CoVs) and low-pathogenicity influenza A viruses (LP IAVs) depend on target cell proteases to cleave their viral glycoproteins and prime them for virus-cell membrane fusion. Several proteases cluster into tetraspanin-enriched microdomains (TEMs), suggesting that TEMs are preferred virus entry portals. Here we found that several CoV receptors and virus-priming proteases were indeed present in TEMs. Isolated TEMs, when mixed with CoV and LP IAV pseudoparticles, cleaved viral fusion proteins to fusion-primed fragments and potentiated viral transductions. That entering viruses utilize TEMs as a protease source was further confirmed using tetraspanin antibodies and tetraspanin short hairpin RNAs ... More