Fig. 6.
PKR-RE1 is involved in DNA-protein interaction.
Electrophoretic mobility shift assay was performed with K562 nuclear extract and oligonucleotide probes designed according to the native core binding motif (PKWT; lanes 1-7) and mutated PKR-RE1 (PKmut; lanes 8-14). Absence (−) and increasing amounts of unlabeled wild-type and mutant competitor are indicated. The figure shows 3 distinct DNA-protein complexes upon incubation with labeled PKWT (lane 1). One of these bands (arrow) could be competed off successfully by increasing amounts of excess unlabeled PKWT (lanes 2-4), but remained unaffected by the addition of increasing amounts of excess unlabeled PKmut as a competitor (lanes 5-7). In contrast, this band was absent when lysates were incubated with radiolabeled PKmut (lane 8), whereas the extra band that appeared upon incubation with PKmut could be competed off succesfully by increasing amounts of both unlabeled PKWT and PKmut (lanes 9-11 and 12-14, respectively).