FIGURE 8.

C-terminal DUE-B mutants inhibit Cdc45 loading in DUE-B-depleted egg extracts. A, His₆-tagged DUE-B C-terminal constructs were expressed in HeLa cells and isolated using Ni-NTA beads. The isolated proteins were added to anti-DUE-B immunodepleted (▵) egg extracts and detected by Western blotting. Mock depleted, mock ▵. B, sperm chromatin was added to extracts shown in panel A. DNA synthesis was monitored by [α-³²P]dCTP incorporation (autoradiogram). C, sperm chromatin was purified from replication reactions and analyzed by Western blotting.