Figure 5
From: Identification of archaeal proteins that affect the exosome function in vitro
Influence of PaNip7 and mutants on the Pa-exosome RNA polyadenylation activity. Polyadenylation reactions were performed with a radiolabeled 14-mer poly-A RNA probe incubated with 100 pmol of PaNip7, PaNip7R151A, R152A, or PaNip7K155A, K158A, and different amounts of the exosome complexes (5 or 10 pmol of RNase PH ring, PaRrp4-exosome, or PaCsl4-exosome). Proteins were incubated with 1 pmol RNA at 37°C for 30 min, in the presence of 10 mM ADP and 10 mM MgCl2. Samples were separated on 8% denaturing polyacrylamide gels and visualized by phosphorimaging. -, No protein added to the reaction. RNA oligo and polymerization products are indicated on the right-hand side.