Fig. 2.

Activation of MACV L by the 5′ ligand is sequence-specific. (A, Upper) Comparison of the MACV conserved terminal 5′ vRNA sequence with La Crosse virus (LACV) and influenza (Flu) RdRP atomic structures. (A, Lower) MACV 5′:3′ v/cRNA duplexes containing S segment sequences represented in GenBank. (B) MACV L is specifically activated by a conserved arenavirus sequence. In vitro RNA synthesis reactions were performed as in Fig. 1. The gel labels for the Middle and Right hand panels correspond to the oligo sequences shown in A and B, respectively. (C) The 5′ vRNA and 5′ cRNA of MACV mediate activation of L, in a template-dependent manner. In vitro RNA synthesis reactions were performed with either a 3′ vRNA template (Left) or a 3′ cRNA template (Right). For each template, a 12-nt 5′ vRNA or 5′ cRNA ligand was included. Products of RNA synthesis were visualized and quantified as in Fig. 1, and the relative changes in RNA accumulation are shown as bar graphs below the gel images in B and C. Error bars for all graphs represent the SD from the mean of three independent experiments.