14-3-3σ is required to prevent mitotic catastrophe after DNA damage

14-3-3 sigma is a member of a family of proteins that regulate cellular activity by binding and sequestering phosphorylated proteins. It has been suggested that 14-3-3 sigma promotes pre-mitotic cell-cycle arrest following DNA damage, and that its expression can be controlled by the p53 tumour suppressor gene(1). Here we describe an improved approach to the generation of human somatic-cell knockouts, which we have used to generate human colorectal cancer cells in which both 14-3-3 sigma alleles are inactivated. After DNA damage, these cells initially arrested in the G2 phase of the cell cycle, but, unlike Cells containing 14-3-3 sigma, the 14-3-3 sigma(-/-) cells were unable to maintain cell-cycle arrest. The 14-3-3 sigma(-/-) cells died ('mitotic catastrophe') as they entered mitosis. This process was associated with a failure of the 14-3-3 sigma-deficient cells to sequester the proteins (cyclin B1 and cdc2) that initiate mitosis and prevent them from entering the nucleus. These results may indicate a mechanism for maintaining the G2 checkpoint and preventing mitotic death.