Parham, J.; Cezanne, A.; Sorichetti, V.; Hoogenberg, B.; Foo, S.; Kuo, Y.-w.; Mawdesley, E.; Gatward, L. D.; Schulze, U.; Boulanger, J.; Saric, A.; Baum, B.

Cells must coordinate DNA segregation with cytokinesis to ensure that each daughter cell inherits a complete genome. Here, we explore how DNA segregation and division are mechanistically coupled in archaeal relatives of eukaryotes, which lack CDK/Cyclins. Using live cell imaging we first describe the series of sequential changes in DNA organisation that accompany cell division in Sulfolobus, which computational modelling shows likely aid genome segregation. Through a perturbation analysis we identify a regulatory checkpoint which ensures that the compaction of the genome into two spatially segregated nucleoids only occurs once cells have assembly a division ring - which also defines the axis of DNA segregation. Finally, we show that DNA compaction and segregation depends, in part, on a ParA homologue, SegA, and its partner SegB, whose absence leads to bridging DNA. Taken together, these data show how regulatory checkpoints like those operating in eukaryotes aid high-fidelity division in an archaeon.