Realtime 2-5A kinetics suggests interferons ß and ? evade global arrest of translation by RNase L

Alisha Chitrakar; Sneha Rath; Jesse Donovan; Kaitlin Demarest; Yize Li; Raghavendra Rao Sridhar; Susan R. Weiss; Sergei V. Kotenko; Ned S. Wingreen; Alexei Korennykh

Toward this end, we engineered tagged versions of V6 with nuclear localization signal (NLS) and nuclear export signal (NES). Both variants localized to the expected sites ( Fig. 3C ). In the presence of poly-IC, NLS-V6 and NES-V6 produced nearly identical luminescence profiles, which are similar to the trace obtained with untagged V6 (Fig. 3D ). This similarity is most simply reconciled with a model of rapid 2-5A equilibration between the two compartments due to diffusion. Indeed, 2-5A produced at the center of a HeLa cell, may take only several seconds to diffuse across the nucleus, through the nuclear pores, and to become evenly distributed between the nucleus and the cytosol ( Fig. 3E ). Our measurements did not support localized 2-5A action and indicated that 2-5A was poised to establish communication between the OASs and RNase L across the cell.