Figure 7.
Analysis of Nsp15-RNA complex. (a) Gel-filtration profile of the catalytically inactive Nsp15 mutant, H249A in the presence (unbroken line) or in the absence (broken line) of biotin-labeled RNA U10. The positions where hexamer, trimer and monomer elute were characterized by Guarino et al.3 (b) Individual gel-filtration fractions on SDS-PAGE. (c) Slot blot of gel filtration fractions obtained when Nsp15 and RNA subjected to UV crosslinking were passed through the column. (d) Slot blot analysis of gel-filtration fractions performed with only RNA. Fraction numbers are indicated above the gel and blots. The RNAs in the blots were detected by probing with horse-radish peroxidase-conjugated streptavidin and developed using enhanced chemiluminiscence.