title: Bub1 autophosphorylation feeds back to regulate kinetochore docking and promote localized substrate phosphorylation.
abstract: During mitosis, Bub1 kinase phosphorylates histone H2A-T120 to promote centromere sister chromatid cohesion through recruitment of shugoshin (Sgo) proteins. The regulation and dynamics of H2A-T120 phosphorylation are poorly understood. Using quantitative phosphoproteomics we show that Bub1 is autophosphorylated at numerous sites. We confirm mitosis-specific autophosphorylation of a several residues and show that Bub1 activation is primed in interphase but fully achieved only in mitosis. Mutation of a single autophosphorylation site T589 alters kinetochore turnover of Bub1 and results in uniform H2A-T120 phosphorylation and Sgo recruitment along chromosome arms. Consequently, improper sister chromatid resolution and chromosome segregation errors are observed. Kinetochore tethering of Bub1-T589A refocuses H2A-T120 phosphorylation and Sgo1 to centromeres. Recruitment of the Bub1-Bub3-BubR1 axis to kinetochores has recently been extensively studied. Our data provide novel insight into the regulation and kinetochore residency of Bub1 and indicate that its localization is dynamic and tightly controlled through feedback autophosphorylation.
authors.affiliation: Faculty of Medicine, Department of Molecular and Cellular Biology, Université Laval, Québec, Canada G1V 0A6.|Department of Reproduction, Mother and Youth Health, Centre de recherche du Centre Hospitalier Universitaire de Québec, Québec, Canada G1V 4G2.
authors.forename: Adeel
authors.initials: A
authors.lastname: Asghar
authors.affiliation: Faculty of Medicine, Department of Molecular and Cellular Biology, Université Laval, Québec, Canada G1V 0A6.
authors.forename: Audrey
authors.initials: A
authors.lastname: Lajeunesse
authors.affiliation: ProQR Therapeutics N.V., Darwinweg 24, Leiden 2333 CR, The Netherlands.
authors.forename: Kalyan
authors.initials: K
authors.lastname: Dulla
authors.affiliation: Faculty of Medicine, Department of Molecular and Cellular Biology, Université Laval, Québec, Canada G1V 0A6.|Department of Reproduction, Mother and Youth Health, Centre de recherche du Centre Hospitalier Universitaire de Québec, Québec, Canada G1V 4G2.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Combes
authors.affiliation: Department of Reproduction, Mother and Youth Health, Centre de recherche du Centre Hospitalier Universitaire de Québec, Québec, Canada G1V 4G2.
authors.forename: Philippe
authors.initials: P
authors.lastname: Thebault
authors.affiliation: Biozentrum, University of Basel, Klingelbergstrasse 50/70, Basel CH-4056, Switzerland.
authors.forename: Erich A
authors.initials: EA
authors.lastname: Nigg
authors.affiliation: Faculty of Medicine, Department of Molecular and Cellular Biology, Université Laval, Québec, Canada G1V 0A6.|Department of Reproduction, Mother and Youth Health, Centre de recherche du Centre Hospitalier Universitaire de Québec, Québec, Canada G1V 4G2.
authors.forename: Sabine
authors.initials: S
authors.lastname: Elowe
mesh_terms: D018797:Cell Cycle Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002503:Centromere / Q000378:metabolism; D002853:Chromatography, Liquid; D025461:Feedback, Physiological; D005455:Fluorescent Antibody Technique; D006367:HeLa Cells; D006657:Histones / Q000378:metabolism*; D006801:Humans; D007399:Interphase; D018386:Kinetochores / Q000378:metabolism*; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D008938:Mitosis; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D000075223:Poly-ADP-Ribose Binding Proteins; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: C489444:BUB3 protein, human; D018797:Cell Cycle Proteins; D006657:Histones; D000075223:Poly-ADP-Ribose Binding Proteins; C496644:SGO1 protein, human; C584277:BUB1 protein, human; D017346:Protein Serine-Threonine Kinases