Figure 4
From: Identification of archaeal proteins that affect the exosome function in vitro
Effect of PaNip7 mutants on the Pa-exosome poly-AU RNA degradation activity. RNase activity reactions were performed by incubating 1 pmol of a radiolabeled 21-mer poly-rAU with 100 pmol of either PaNip7, PaNip7R151A, R152A, or PaNip7K155A, K158A, for 30 min at 37°C, after which time increasing amounts of the exosome complexes (1, 5 or 10 pmol of either RNase PH ring, PaRrp4-exosome, or PaCsl4-exosome), and NaH2PO4 (10 mM) were added to the reactions, and further incubated for 15 min at 65°C. Samples were separated on 8% denaturing polyacrylamide gels and visualized by phosphorimaging. -, No protein added to the reaction. RNA oligo and degradation products are indicated on the right-hand side.