Separase is required at multiple pre-anaphase cell cycle stages in human cells.
The yeast separase proteins Esp1 and Cut1 are required for loss of sister chromatid cohesion that occurs at the moment of anaphase onset. Circumstantial evidence has linked human separase to centromere separation at anaphase, but a direct test that the role of this enzyme is functionally conserved with the yeast proteins is lacking. Here we describe the effects of separase depletion from human cells using RNA interference. Surprisingly, HeLa cells lacking separase are delayed or arrest at the G2-M phase transition. This arrest is not likely due to the activation of a known checkpoint control, but may be a result of a failure to construct a mitotic chromosome. Without separase, cells also have a prolonged prometaphase, perhaps resulting from defects in spindle assembly or dynamics. In cells that reach mitosis, sister arm resolution and separation are perturbed, whereas in anaphase cells sister centromeres do appear to separate. These data indicate that separase function is not restricted to anaphase initiation and that its role in promoting loss of sister chromatid cohesion might be preferentially at arms but not centromeres.
Giménez-Abián
Juan F
JF
Department of Genetics, Cell Biology & Development, University of Minnesota Medical School, Minneapolis, Minnesota 55455, USA.
Díaz-Martínez
Laura A
LA
Waizenegger
Irene C
IC
Giménez-Martín
Gonzalo
G
Clarke
Duncan J
DJ
D002453:Cell Cycle / Q000235:genetics / Q000502:physiology*; D018797:Cell Cycle Proteins / Q000235:genetics / Q000502:physiology*; D002455:Cell Division / Q000235:genetics; D002503:Centromere / Q000737:chemistry / Q000235:genetics; D010450:Endopeptidases / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D016195:G2 Phase / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D034622:RNA Interference; D064247:Separase
D018797:Cell Cycle Proteins; D010450:Endopeptidases; C577987:ESPL1 protein, human; D064247:Separase