title: WT1 interacts with MAD2 and regulates mitotic checkpoint function.
abstract: Tumour suppressors safeguard the fidelity of the mitotic checkpoint by transcriptional regulation of genes that encode components of the mitotic checkpoint complex (MCC). Here we report a new role for the tumour suppressor and transcription factor, WT1, in the mitotic checkpoint. We show that WT1 regulates the MCC by directly interacting with the spindle assembly checkpoint protein, MAD2. WT1 colocalizes with MAD2 during mitosis and preferentially binds to the functionally active, closed-conformer, C-MAD2. Furthermore, WT1 associates with the MCC containing MAD2, BUBR1 and CDC20, resulting in prolonged inhibition of the anaphase-promoting complex/cyclosome (APC/C) and delayed degradation of its substrates SECURIN and CYCLIN B1. Strikingly, RNA interference-mediated depletion of WT1 leads to enhanced turnover of SECURIN, decreased lag time to anaphase and defects in chromosome segregation. Our findings identify WT1 as a regulator of the mitotic checkpoint and chromosomal stability.
authors.affiliation: Department of Biological Sciences, University at Buffalo, Cooke Hall, North Campus, Buffalo, New York 14260, USA.
authors.forename: Jayasha
authors.initials: J
authors.lastname: Shandilya
authors.affiliation: Department of Biological Sciences, University at Buffalo, Cooke Hall, North Campus, Buffalo, New York 14260, USA.
authors.forename: Eneda
authors.initials: E
authors.lastname: Toska
authors.affiliation: School of Biomedical Sciences, Institute of Health and Biomedical Innovation at the Translational Research Institute, Queensland University of Technology, Brisbane, Queensland 4102, Australia.
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Richard
authors.affiliation: Department of Biological Sciences, University at Buffalo, Cooke Hall, North Campus, Buffalo, New York 14260, USA.
authors.forename: Kathryn F
authors.initials: KF
authors.lastname: Medler
authors.affiliation: 1] Department of Biological Sciences, University at Buffalo, Cooke Hall, North Campus, Buffalo, New York 14260, USA [2] School of Cellular and Molecular Medicine, University of Bristol, Bristol BS8 1TD, UK.
authors.forename: Stefan G E
authors.initials: SG
authors.lastname: Roberts
mesh_terms: D000818:Animals; D064199:Cdc20 Proteins / Q000378:metabolism; D045744:Cell Line, Tumor; D002875:Chromosomes / Q000737:chemistry / Q000648:ultrastructure; D056744:Cyclin B1 / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D005982:Glutathione Transferase / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D020014:K562 Cells; D061986:MCF-7 Cells; D064246:Mad2 Proteins / Q000378:metabolism*; D051379:Mice; D008938:Mitosis; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism; D017434:Protein Structure, Tertiary; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D064249:Securin / Q000378:metabolism; D020798:Two-Hybrid System Techniques; D025721:WT1 Proteins / Q000378:metabolism*
chemical_list: C534699:CCNB1 protein, human; D064199:Cdc20 Proteins; D056744:Cyclin B1; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D034741:RNA, Small Interfering; D064249:Securin; D025721:WT1 Proteins; C583751:WT1 protein, human; C578002:pituitary tumor-transforming protein 1, human; C577488:CDC20 protein, human; D005982:Glutathione Transferase; C584277:BUB1 protein, human; D017346:Protein Serine-Threonine Kinases