Separase biosensor reveals that cohesin cleavage timing depends on phosphatase PP2A(Cdc55) regulation.
In anaphase, sister chromatids separate abruptly and are then segregated by the mitotic spindle. The protease separase triggers sister separation by cleaving the Scc1/Mcd1 subunit of the cohesin ring that holds sisters together. Polo-kinase phosphorylation of Scc1 promotes its cleavage, but the underlying regulatory circuits are unclear. We developed a separase biosensor in Saccharomyces cerevisiae that provides a quantitative indicator of cohesin cleavage in single cells. Separase is abruptly activated and cleaves most cohesin within 1 min, after which anaphase begins. Cohesin near centromeres and telomeres is cleaved at the same rate and time. Protein phosphatase PP2A(Cdc55) inhibits cohesin cleavage by counteracting polo-kinase phosphorylation of Scc1. In early anaphase, the previously described separase inhibition of PP2A(Cdc55) promotes cohesin cleavage. Thus, separase acts directly on Scc1 and also indirectly, through inhibition of PP2A(Cdc55), to stimulate cohesin cleavage, providing a feedforward loop that may contribute to a robust and timely anaphase.
Department of Physiology, University of California-San Francisco, CA 94158, USA.
Gilad
G
Yaakov
Kurt
K
Thorn
David O
DO
Morgan
D015374:Biosensing Techniques; D018797:Cell Cycle Proteins / Q000235:genetics* / Q000378:metabolism*; D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics* / Q000378:metabolism*; D020090:Chromosome Segregation / Q000502:physiology*; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008856:Microscopy, Fluorescence / Q000379:methods; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics* / Q000378:metabolism; D064247:Separase; D000097722:Cohesins
D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D010450:Endopeptidases; D049452:Green Fluorescent Proteins; D054648:Protein Phosphatase 2; D029701:Saccharomyces cerevisiae Proteins; D064247:Separase; D000097722:Cohesins; C071246:CDC55 protein, S cerevisiae; C490529:MCD1 protein, S cerevisiae; C577986:ESP1 protein, S cerevisiae