The Drosophila Cdc6/18 protein has functions in both early and late S phase in S2 cells

The Cdc6/18 protein has been mainly characterised for its role in the initiation of DNA replication. Several studies exist, however, which suggest that it may also have a role in controlling the G2/M transition. Here we present studies on the Drosophila Cdc6 (DmCdc6) protein that support this dual function for the protein. First we show that its location is consistent with a cellular role post replication initiation as it remains nuclear throughout G1, S and G2 phases. In addition, we have been able to reduce the level of DmCdc6 protein to nondetectable levels in S2 cells using RNAi. This causes DNA fragmentation and cell cycle abnormalities which have some similarities with phenotypes previously observed in yeasts and are consistent with the cells entering mitosis with incompletely replicated DNA. Finally, we have stably overexpressed the DmCdc6 protein to a high level in S2 cells. Despite a large excess of protein the effects on the S2 cells were minimal. We did, however, detect a slight stalling of the cells in the late S phase of the cell cycle, which further supports the proposal that DmCdc6 has a role in controlling the transition from the S to M phases of the cycle.