Figure 3.

ATP requirement for protein assembly on ARS1 beads. (A) ARS 1 beads were incubated with whole-cell extracts prepared from YGP82 cells expressing Cdc6p either in the presence of (ATP +) or in the absence of (ATP −) exogenously added ATP and the ATP regenerating system. Proteins associating with the beads (Bead) or remaining in the supernatants (Sup) were detected by immunoblotting as before. (B) Loading reactions were carried out either with or without exogenously added ATP and the ATP regenerating system (ATP + or −). To the indicated reactions (Apyrase +), 5 units of apyrase was added. After incubation with ARS1 beads, Orc2p associating with the beads (Bead) and remaining in the supernatants (Sup) was detected.