Figure 4.
Effect of GST–FFA-1 fusion protein on DNA replication. (A) Replication of nuclei reconstituted around sperm chromatin in the presence of various fusion proteins (500 nM final concentration for GST-Xho and GST-Stu/Xho; 1 μM for GST-Stu and GST) or buffer. The fusion proteins and [32P]dATP were added at the beginning of the reactions. Samples were taken at the indicated times and separated on 0.8% agarose/TAE gel. The top band is the DNA that was too large to enter the gel and the bottom band is the sheared DNA (>20 kb) introduced during sample preparation. (B) Diagram of the GST–FFA-1 fusion proteins used in this study. Numbers indicate the positions of the amino acids that demarcate the various domains in FFA-1. (C) The fusion proteins used in this study separated on a 12% SDS polyacrylamide gel and stained by Coomassie blue. Molecular weight markers are labeled on the right in kilodaltons.