Figure 4.

Determination of the region of AlF-C2 that binds to Orc1. (A) Various deletion mutants of AlF-C2, schematically presented in (B), were prepared as GST-tagged proteins and bound to glutathione beads. T7-tagged Orc1 and Orc2 were incubated with glutathione beads that were coupled with GST–AlF-C2 deletion mutants. Proteins bound to the beads were separated by SDS–PAGE and blotted with anti-T7 antibody (upper panel). The 1/10 volume of unbound fractions was also analyzed (lower panel). (B) Schematic representation of AlF-C2 and its truncation mutants. The RRM and Orc1-binding domain are shown with amino acid residue number.