Figure 5

Orf1-RNA binding activity. (A) Schematic organization of the abiD1 gene. Arrows indicate position of abiD1 transcripts (1 to 7) used for binding experiments. (B) Binding of purified Orf1, OrfM1 and Orf1M2 to the abiD1 mRNA 1. Ten ng of radiolabelled abiD1 mRNA (transcript 1) were incubated with increasing amounts of Orf1, OrfM1 or OrfM2 protein (0; 0.03 μM; 0.05 μM; 0.11 μM; 0.22 μM), followed by separation of the nucleoprotein complex in an 8% non-denaturing polyacrylamide gel at room temperature. (C) Binding of Orf1 to the abiD1 mRNA 6 and abiD1 mRNA 7. (D) Binding of Orf1 to trpA mRNA and aldB mRNA. Experiments (C and D) were performed with increasing amounts of Orf1 protein (0; 0.11 μM; 0.22 μM; 0.33 μM) in the same conditions as above.