Evolutionary dynamics of the kinetochore network in eukaryotes as revealed by comparative genomics

Hooff, Jolien J. E. van; Tromer, Eelco C.; Wijk, Leny M. van; Snel, Berend; Kops, Geert J. P. L.

doi:10.15252/embr.201744102

Cited by 223 publications

(405 citation statements)

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“…Given that our quantitative analysis of Bub1 CR and Rod CR cells indicated an integrated function of the two checkpoint components, we hypothesized that RZZ localizes Mad1 on kinetochores to facilitate the interaction between Bub1 and Mad1. Interestingly, plants and algae lack the RZZ complex and one of their three Bub1 like proteins contains multiple repeats of the CD1 domain likely to increase the strength of the Mad1-Bub1 interaction (Di Fiore et al, 2016;van Hooff et al, 2017). In this assay, Mad1 is fused to the BirA* biotin ligase resulting in biotinylation of proteins in proximity of Mad1 upon addition of biotin to the cell culture.…”

Section: Increasing the Interaction Between Bub1 And Mad1 Bypasses Thmentioning

confidence: 99%

Efficient mitotic checkpoint signaling depends on integrated activities of Bub1 and the RZZ complex

et al. 2019

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Kinetochore localized Mad1 is essential for generating a “wait anaphase” signal during mitosis, hereby ensuring accurate chromosome segregation. Inconsistent models for the function and quantitative contribution of the two mammalian Mad1 kinetochore receptors: Bub1 and the Rod‐Zw10‐Zwilch (RZZ) complex exist. By combining genome editing and RNAi, we achieve penetrant removal of Bub1 and Rod in human cells, which reveals that efficient checkpoint signaling depends on the integrated activities of these proteins. Rod removal reduces the proximity of Bub1 and Mad1, and we can bypass the requirement for Rod by tethering Mad1 to kinetochores or increasing the strength of the Bub1‐Mad1 interaction. We find that Bub1 has checkpoint functions independent of Mad1 localization that are supported by low levels of Bub1 suggesting a catalytic function. In conclusion, our results support an integrated model for the Mad1 receptors in which the primary role of RZZ is to localize Mad1 at kinetochores to generate the Mad1‐Bub1 complex.

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Section: Increasing the Interaction Between Bub1 And Mad1 Bypasses Thmentioning

confidence: 99%

Efficient mitotic checkpoint signaling depends on integrated activities of Bub1 and the RZZ complex

et al. 2019

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“…The RZZ complex is conserved in metazoans and other taxa [18,35,36], but its roles in SAC signaling and kinetochore organization remain poorly understood. Our studies define RZZ as a critical SAC maintenance factor, the major Mad1-Mad2 tether at kinetochores, and a key mediator of phosphodependent kinetochore expansion.…”

Section: Discussionmentioning

confidence: 99%

The RZZ complex integrates spindle checkpoint maintenance with dynamic expansion of unattached kinetochores

Rodriguez-Rodriguez

McKinley

Sikirzhytski

et al. 2018

Preprint

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SummaryThe Mad1-Mad2 heterodimer is the catalytic hub of the spindle assembly checkpoint (SAC), which controls mitosis through assembly of a multi-subunit anaphase inhibitor, the mitotic checkpoint complex (MCC) [1,2]. Mad1-Mad2 first catalyzes MCC assembly at interphase nuclear pores [3], then migrates to kinetochores at nuclear envelope breakdown (NEBD) and resumes MCC assembly until bipolar spindle attachment is complete [1,2]. There is significant debate about the factor(s) involved in targeting Mad1-Mad2 to kinetochores in higher eukaryotes [4][5][6][7][8][9]. Through gene editing and livecell imaging, we found that the human Rod-Zw10-Zwilch (RZZ) complex is dispensable for cell viability and initial recruitment of Mad1-Mad2 to kinetochores at NEBD, but then becomes necessary to tether Mad1-Mad2 at kinetochores and sustain SAC arrest in cells challenged with spindle poisons. We also show that RZZ forms the mesh-like fibrous corona, a structural expansion of the outer kinetochore important for timely chromosome congression [10][11][12][13] once Mps1 phosphorylates the N-terminus of Rod.Artificially tethering Mad1-Mad2 to kinetochores enabled long-term mitotic arrest in the absence of RZZ. Conversely, blocking early RZZ-independent recruitment of Mad1-Mad2 eliminated the transient SAC response in RZZ-null cells. We conclude that RZZ drives structural changes in the outer kinetochore that facilitate chromosome biorientation and chronic SAC transduction, a key determinant of cytotoxicity during antimitotic drug therapy [14][15][16].All rights reserved. No reuse allowed without permission.was not peer-reviewed) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity.The copyright holder for this preprint (which . http://dx.doi.org/10.1101/297580 doi: bioRxiv preprint first posted online Apr. 9, 2018; 3 Results RZZ is required for long-term mitotic arrest in response to spindle poisonsRecent studies have reached different conclusions about the specific roles and relative importance of Bub1 and the RZZ complex in targeting Mad1-Mad2 to kinetochores and transducing SAC arrest [4][5][6][7][8]. To interrogate RZZ function with maximum penetrance, we used AAV (adeno-associated virus)-and CRISPR-mediated gene editing to target the KNTC1 (Rod) locus in HCT116 cells, a diploid colorectal cell line ( Fig. S1A-C). The resulting KNTC1 HF/-(hypomorph-flox) cells expressed Rod at ~20% of the wildtype level ( Fig. 1A-B and S1D) and exited mitosis prematurely when microtubule polymerization (nocodazole, 99 ± 6 min s.e.m.) or Eg5-dependent spindle bipolarity (S-trityl-L-cysteine (STLC), 193 ± 9 min s.e.m.) were inhibited, whereas wildtype cells never exited mitosis during the 16-hour timelapse (Fig. 1A-B). To determine if complete loss of the RZZ complex is compatible with clonogenic survival, KNTC1 HF/-cells were transduced with an adenovirus expressing Cre recombinase (AdCre) and subjected to limiting dilution.Unlike most SAC gene knockouts in mammals, KNTC1 -/-cells were viable (Fig. 1...

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“…Zwint‐1 (human), Kre28 (budding yeast), and Sos7 (fission yeast) are orthologous to one another and belong to a single OG. This conclusion was based on successive similarity searches that indicated homology between various Zwint‐1‐like, Kre28‐like, and Sos7‐like proteins of different fungal, animal, and animal‐related species . Starting with the Sos7 protein (fission yeast, Schizosaccharomyces pombe ), the ordered searches indicate which “bridging” sequences were used to establish orthology to Kre28 and Zwint‐1.…”

Section: Challenging the Evolutionary Scenariomentioning

confidence: 99%

“…Due to their intimate knowledge of proteins and cellular processes, these biologists can provide invaluable insights into evolutionary interpretations posed by evolutionary biologists. In collaboration with molecular biologists, we studied diverse eukaryotic proteins, varying from chromatin remodelers and bZIP transcription factors to flagellum and kinetochore components . We experienced that for non‐bioinformaticians various challenges arise regarding the use of tools, the knowledge of genome evolution and of the species tree, and, importantly, how complicated evolutionary histories are reflected in a myriad of computational tools and sequence databases.…”

Section: Introductionmentioning

confidence: 99%

“…In collaboration with molecular biologists, we studied diverse eukaryotic proteins, varying from chromatin remodelers and bZIP transcription factors to flagellum and kinetochore components. [2,[33][34][35] We experienced that for nonbioinformaticians various challenges arise regarding the use of tools, the knowledge of genome evolution and of the species tree, and, importantly, how complicated evolutionary histories are reflected in a myriad of computational tools and sequence databases. Such challenges may cause flawed evolutionary inferences, which in turn may cause incorrect, or incomplete, function prediction.…”

Section: Introductionmentioning

confidence: 99%

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Inferring the Evolutionary History of Your Favorite Protein: A Guide for Molecular Biologists

et al. 2019

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Comparative genomics has proven a fruitful approach to acquire many functional and evolutionary insights into core cellular processes. Here it is argued that in order to perform accurate and interesting comparative genomics, one first and foremost has to be able to recognize, postulate, and revise different evolutionary scenarios. After all, these studies lack a simple protocol, due to different proteins having different evolutionary dynamics and demanding different approaches. The authors here discuss this challenge from a practical (what are the observations?) and conceptual (how do these indicate a specific evolutionary scenario?) viewpoint, with the aim to guide investigators who want to analyze the evolution of their protein(s) of interest. By sharing how the authors draft, test, and update such a scenario and how it directs their investigations, the authors hope to illuminate how to execute molecular evolution studies and how to interpret them. Also see the video abstract here https://youtu.be/VCt3l2pbdbQ.

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Evolutionary dynamics of the kinetochore network in eukaryotes as revealed by comparative genomics

Cited by 223 publications

References 96 publications

Efficient mitotic checkpoint signaling depends on integrated activities of Bub1 and the RZZ complex

Efficient mitotic checkpoint signaling depends on integrated activities of Bub1 and the RZZ complex

The RZZ complex integrates spindle checkpoint maintenance with dynamic expansion of unattached kinetochores

Inferring the Evolutionary History of Your Favorite Protein: A Guide for Molecular Biologists

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