Negative feedback at kinetochores underlies a responsive spindle checkpoint signal.
Kinetochores are specialized multi-protein complexes that play a crucial role in maintaining genome stability. They bridge attachments between chromosomes and microtubules during mitosis and they activate the spindle assembly checkpoint (SAC) to arrest division until all chromosomes are attached. Kinetochores are able to efficiently integrate these two processes because they can rapidly respond to changes in microtubule occupancy by switching localized SAC signalling ON or OFF. We show that this responsiveness arises because the SAC primes kinetochore phosphatases to induce negative feedback and silence its own signal. Active SAC signalling recruits PP2A-B56 to kinetochores where it antagonizes Aurora B to promote PP1 recruitment. PP1 in turn silences the SAC and delocalizes PP2A-B56. Preventing or bypassing key regulatory steps demonstrates that this spatiotemporal control of phosphatase feedback underlies rapid signal switching at the kinetochore by: allowing the SAC to quickly transition to the ON state in the absence of antagonizing phosphatase activity; and ensuring phosphatases are then primed to rapidly switch the SAC signal OFF when kinetochore kinase activities are diminished by force-producing microtubule attachments.
1] Department of Medical Oncology, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [2] Molecular Cancer Research, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [3] Center for Molecular Medicine, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands.
Wilco
W
Nijenhuis
Division of Cancer Research, Medical Research Institute, Jacqui Wood Cancer Centre, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, UK.
Giulia
G
Vallardi
1] Department of Medical Oncology, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [2] Molecular Cancer Research, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [3] Center for Molecular Medicine, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands.
Antoinette
A
Teixeira
1] Department of Medical Oncology, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [2] Molecular Cancer Research, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [3] Center for Molecular Medicine, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands [4] Cancer Genomics Netherlands, UMC Utrecht, Universiteitsweg 100, 3584CG Utrecht, The Netherlands.
Geert J P L
GJ
Kops
Division of Cancer Research, Medical Research Institute, Jacqui Wood Cancer Centre, Ninewells Hospital and Medical School, University of Dundee, Dundee DD1 9SY, UK.
Adrian T
AT
Saurin
D020816:Amino Acid Motifs / Q000235:genetics; D064107:Aurora Kinase B / Q000037:antagonists & inhibitors; D018797:Cell Cycle Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D045744:Cell Line, Tumor; D020090:Chromosome Segregation; D025461:Feedback, Physiological; D006367:HeLa Cells; D006801:Humans; D018386:Kinetochores / Q000378:metabolism*; D059566:M Phase Cell Cycle Checkpoints / Q000235:genetics*; D008869:Microtubule-Associated Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D008870:Microtubules / Q000378:metabolism; D015739:Nocodazole / Q000494:pharmacology; D010766:Phosphorylation; D011485:Protein Binding; D054648:Protein Phosphatase 2 / Q000235:genetics / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics; D034622:RNA Interference; D034741:RNA, Small Interfering; D015398:Signal Transduction; D051899:Smad2 Protein / Q000235:genetics; D008941:Spindle Apparatus / Q000235:genetics*; D050257:Tubulin Modulators / Q000494:pharmacology
D018797:Cell Cycle Proteins; C472643:Knl1 protein, human; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; C496644:SGO1 protein, human; C497782:SMAD2 protein, human; D051899:Smad2 Protein; D050257:Tubulin Modulators; D011505:Protein-Tyrosine Kinases; C576921:AURKB protein, human; D064107:Aurora Kinase B; C584277:BUB1 protein, human; D017346:Protein Serine-Threonine Kinases; C075840:TTK protein, human; C572369:PP2A-B56alpha protein, human; D054648:Protein Phosphatase 2; D015739:Nocodazole