The N-Terminal Domain of the Arenavirus L Protein Is an RNA Endonuclease Essential in mRNA Transcription
Figure 3
Nucleotide and RNA binding assays of LCMV NL1 domain.
A, Cross-linking assay. 7 µg of purified protein were incubated in the absence (−) or presence of each indicated radiolabelled NTP. The mixture was then UV-irradiated and loaded onto a denaturing polyacrylamide gel. The latter was analyzed by autoradiography (top) and Coomassie blue staining (bottom). B, Band shift assay. Radiolabelled RNA was incubated with increasing quantities (1.4 µg (+), 4.2 µg (++) and 7 µg (+++)) of NL1 protein. Reaction mixture was then analyzed by PAGE, and the gel was visualized by autoradiography (left) and Coomassie blue staining (right). Apparent degradation products are indicated by arrows under the RNA input arrow.