Fig. P1.

(A) Rabies viral lifecycle. (B) Cartoon of the CFPS system, which can faithfully reconstitute intracellular biochemical interactions in vitro. (C) Model for interaction of newly synthesized rabies proteins with host multiprotein assembly complex, as reconstituted by CFPS. Colored shapes represent host factors; black shapes represent viral proteins. The host complex shown here is a member of a hypothesized set of assembly machines in the cytosol composed of multiple different polypeptides with accessory components recruited or lost depending on the substrate bound. The assembly machine is shown binding rabies phosphoprotein (P) and nascent rabies nucleoprotein (N) and progressing through a series of assembly intermediates. Note the changing orientation/configuration of the assembly machine. PAV-866 is hypothesized to bind an allosteric site on the assembly machine to disrupt interactions effectively, resulting in the formation of aberrant, off-pathway complexes.