Sister chromatid cohesion and separation are fundamental for accurate genome inheritance over cell generations. Work over recent years has established the existence of a chromosomal protein complex, cohesin, that connects sister chromatids from the time they are generated in S phase onwards, and which is destroyed at the onset of anaphase through cleavage by the protease separase. Over the last year, the function of cohesin has been investigated in higher eukaryotes, including humans, with results that have uncovered important new aspects of this process. The first structural views of cohesin have become available, and significant steps been made towards a mechanistic understanding of chromosome cohesion. Studies on separase have revealed new levels of regulation of chromosome segregation.