Cell Cycle-dependent Subcellular Translocation of the Human DNA Licensing Inhibitor Geminin

Once per cell cycle replication is crucial for maintaining genome integrity. Geminin interacts with the licensing factor Cdt1 to prevent untimely replication and is controlled by APC/C-dependent cell cycle specific proteolysis during mitosis and in G 1 . We show here that human geminin, when expressed in human cells in culture under a constitutive promoter, is excluded from the nucleus during part of the G 1 phase and at the transition from G 0 to G 1 . The N-terminal 30 amino acids of geminin, which contain its destruction box, are essential for nuclear exclusion. In addition, 30 amino acids within the central domain of geminin are required for both nuclear exclusion and nuclear accumulation. Cdt1 overexpression targets geminin to the nucleus, while reducing Cdt1 levels by RNAi leads to the appearance of endogenous geminin in the cytoplasm. Our data propose a novel means of regulating the balance of Cdt1/geminin in human cells, at the level of the subcellular localization of geminin. Background: The licensing inhibitor geminin must be regulated to ensure once per cell cycle replication. Results: Geminin is spatially and temporally controlled by nuclear exclusion during part of the G 1 phase in human cells. Conclusion: Geminin exclusion from the nucleus provides an additional level of licensing control, balancing the Cdt1/geminin ratio. Significance: Multiple overlapping mechanisms are used by human cells to ensure genome stability.