Cell-Free, De Novo Synthesis of Poliovirus

TLDR

The study aims to enable cell‑free synthesis of poliovirus to facilitate research on picornavirus replication and control. Poliovirus RNA was translated in a cell‑free HeLa extract, producing viral proteins via polyprotein proteolysis. The cell‑free system produced infectious poliovirus de novo, with virions neutralizable by antiserum and infection blocked by receptor antibodies; synthesis was enhanced 70‑fold by nucleoside triphosphates and inhibited by translation or replication blockers.

Abstract

Cell-free translation of poliovirus RNA in an extract of uninfected human (HeLa) cells yielded viral proteins through proteolysis of the polyprotein. In the extract, newly synthesized proteins catalyzed poliovirus-specific RNA synthesis, and formed infectious poliovirus de novo. Newly formed virions were neutralized by type-specific antiserum, and infection of human cells with them was prevented by poliovirus receptor-specific antibodies. Poliovirus synthesis was increased nearly 70-fold when nucleoside triphosphates were added, but it was abolished in the presence of inhibitors of translation or viral genome replication. The ability to conduct cell-free synthesis of poliovirus will aid in the study of picornavirus proliferation and in the search for the control of picornaviral disease.