dc.contributor.author | Rodriguez-Rodriguez, Jose-Antonio | |
dc.contributor.author | Lewis, Clare | |
dc.contributor.author | McKinley, Kara Lavidge | |
dc.contributor.author | Sikirzhytski, Vitali | |
dc.contributor.author | Corona, Jennifer | |
dc.contributor.author | Maciejowski, John | |
dc.contributor.author | Khodjakov, Alexey | |
dc.contributor.author | Cheeseman, Iain M | |
dc.contributor.author | Jallepalli, Prasad V. | |
dc.date.accessioned | 2020-07-09T14:13:50Z | |
dc.date.available | 2020-07-09T14:13:50Z | |
dc.date.issued | 2018-11 | |
dc.date.submitted | 2018-08 | |
dc.identifier.issn | 0960-9822 | |
dc.identifier.uri | https://hdl.handle.net/1721.1/126113 | |
dc.description.abstract | The Mad1-Mad2 heterodimer is the catalytic hub of the spindle assembly checkpoint (SAC), which controls M phase progression through a multi-subunit anaphase inhibitor, the mitotic checkpoint complex (MCC) [1, 2]. During interphase, Mad1-Mad2 generates MCC at nuclear pores [3]. After nuclear envelope breakdown (NEBD), kinetochore-associated Mad1-Mad2 catalyzes MCC assembly until all chromosomes achieve bipolar attachment [1, 2]. Mad1-Mad2 and other factors are also incorporated into the fibrous corona, a phospho-dependent expansion of the outer kinetochore that precedes microtubule attachment [4–6]. The factor(s) involved in targeting Mad1-Mad2 to kinetochores in higher eukaryotes remain controversial [7–12], and the specific phosphorylation event(s) that trigger corona formation remain elusive [5, 13]. We used genome editing to eliminate Bub1, KNL1, and the Rod-Zw10-Zwilch (RZZ) complex in human cells. We show that RZZ's sole role in SAC activation is to tether Mad1-Mad2 to kinetochores. Separately, Mps1 kinase triggers fibrous corona formation by phosphorylating two N-terminal sites on Rod. In contrast, Bub1 and KNL1 activate kinetochore-bound Mad1-Mad2 to produce a “wait anaphase” signal but are not required for corona formation. We also show that clonal lines isolated after BUB1 disruption recover Bub1 expression and SAC function through nonsense-associated alternative splicing (NAS). Our study reveals a fundamental division of labor in the mammalian SAC and highlights a transcriptional response to nonsense mutations that can reduce or eliminate penetrance in genome editing experiments. Rodriguez-Rodriguez et al. identify distinct roles for Bub1, KNL1, and RZZ in SAC signaling and fibrous corona formation. They also show that BUB1-disrupted clones re-express Bub1 and regain SAC function via nonsense-associated alternative splicing, an often-overlooked transcriptional response that can limit penetrance in genome editing experiments. | en_US |
dc.description.sponsorship | National Institutes of Health (Grant R01GM094972) | en_US |
dc.description.sponsorship | National Institutes of Health (Grant P30CA008748) | en_US |
dc.description.sponsorship | National Institutes of Health (Grant R01GM059363) | en_US |
dc.description.sponsorship | National Institutes of Health (Grant R35GM126930) | en_US |
dc.language.iso | en | |
dc.publisher | Elsevier BV | en_US |
dc.relation.isversionof | http://dx.doi.org/10.1016/j.cub.2018.10.006 | en_US |
dc.rights | Creative Commons Attribution-NonCommercial-NoDerivs License | en_US |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-nd/4.0/ | en_US |
dc.source | PMC | en_US |
dc.title | Distinct Roles of RZZ and Bub1-KNL1 in Mitotic Checkpoint Signaling and Kinetochore Expansion | en_US |
dc.type | Article | en_US |
dc.identifier.citation | Rodriguez-Rodriguez, Jose-Antonio et al. "Distinct Roles of RZZ and Bub1-KNL1 in Mitotic Checkpoint Signaling and Kinetochore Expansion." Current Biology 28, 21 (November 2018): P3422-3429.e5 © 2018 Elsevier | en_US |
dc.contributor.department | Massachusetts Institute of Technology. Department of Biology | en_US |
dc.relation.journal | Current Biology | en_US |
dc.eprint.version | Author's final manuscript | en_US |
dc.type.uri | http://purl.org/eprint/type/JournalArticle | en_US |
eprint.status | http://purl.org/eprint/status/PeerReviewed | en_US |
dc.date.updated | 2019-12-02T15:11:13Z | |
dspace.date.submission | 2019-12-02T15:11:15Z | |
mit.journal.volume | 28 | en_US |
mit.journal.issue | 21 | en_US |
mit.metadata.status | Complete | |