Protein phosphatase 1 regulates exit from the spindle checkpoint in budding yeast.
Accurate chromosome segregation depends on sister kinetochores coming under tension when they make bioriented attachments to microtubules from opposite poles. The spindle checkpoint halts the cell cycle in response to defects in generating proper attachments or tension on kinetochores, although the precise signal that triggers the checkpoint is unclear because tension and attachment are coupled. The target of the checkpoint is the Cdc20 protein, which initiates the anaphase-promoting complex (APC)-dependent degradation of the anaphase inhibitor Pds1/securin. Although the molecular details of spindle checkpoint activation are still being elucidated, phosphorylation by at least four kinases is a crucial requirement. However, less is known about the mechanisms that silence the checkpoint after kinetochores biorient. Here, we show that the catalytic subunit of the budding yeast protein phosphatase 1 (PP1) homolog, Glc7, regulates exit from the checkpoint. Glc7 overexpression prevents spindle checkpoint activation in response to both tension and attachment defects. Although glc7 mutant cells are able to efficiently release from a non-checkpoint-mediated metaphase arrest, they are uniquely sensitive to transient spindle checkpoint activation as a result of a failure in spindle checkpoint exit. We therefore propose that PP1 activity silences the checkpoint by reversing key phosphorylation events.
Pinsky
Benjamin A
BA
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, 1100 Fairview Avenue North, PO Box 19024, Seattle, WA 98109, USA.
Nelson
Christian R
CR
Biggins
Sue
S
D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins / Q000378:metabolism; D020090:Chromosome Segregation / Q000502:physiology*; D017931:DNA Primers / Q000235:genetics; D018816:Genes, cdc / Q000502:physiology*; D015151:Immunoblotting; D018386:Kinetochores / Q000378:metabolism*; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D054645:Protein Phosphatase 1 / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D004718:Saccharomycetales; D008941:Spindle Apparatus / Q000378:metabolism*
C577495:CDC20 protein, S cerevisiae; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins; D017931:DNA Primers; D029701:Saccharomyces cerevisiae Proteins; C092048:GLC7 protein, S cerevisiae; D054645:Protein Phosphatase 1