BubR1 phosphorylates CENP-E as a switch enabling the transition from lateral association to end-on capture of spindle microtubules

Huang, Yunpeng; Lin, Lin; Liu, Xing; Ye, Sheng; Yao, Phil; Wang, Wenwen; Yang, Fengrui; Gao, Xiaolan; Li, Junying; Zhang, Yin; Zhang, Jiancun; Yang, Zhihong; Liu, Xu; Yang, Ziqing; Zang, Jianye; Teng, Maikun; Wang, Zhiyong; Ruan, Ke; Ding, Xia; Li, Lin; Cleveland, Don W.; Zhang, Rongguang; Yao, Xuebiao

doi:10.1038/s41422-019-0178-z

Cited by 56 publications

(66 citation statements)

References 63 publications

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“…Notably, this regulation of BUB1 activity by the BUBR1 pseudokinase may also explain the kinase activity reported in BUBR1 immunoprecipitates, and why this activity is sensitive to mutation of BUBR1 catalytic triad residues, or loss of the BUBR1 kinase which we have shown here tempers BUB1 autophosphorylation 41–43, 73 . In agreement with this, BUB1 and BUBR1 form relatively stable complexes in vitro 44 , and previous work has shown that BUBR1-associated kinase activity is particularly sensitive to BUB1 inhibitors 38 .…”

Section: Discussionsupporting

confidence: 62%

“…The finding that CENP-E played no discernable role in B56 recruitment and SAC silencing was surprising, considering the effect of its inhibition on BUBR1 hyperphosphorylation documented by us and others 41–43, 73 . However, when we examined KARD phosphorylation in CENP-E depleted cells further, we found that while S676 phosphorylation was reduced, as we have shown before, S670 phosphorylation was increased ( 52 and Sup Fig 5A-B ).…”

Section: Resultsmentioning

confidence: 75%

“…Whether BUBR1 is a bona fide active kinase is a subject that has generated considerable controversy 38, 41–43, 73, 78 . If BUBR1 is indeed inactive as proposed before and as we confirm here, an important question that has not yet been addressed is whether the BUBR1 pseudokinase domain retained some function of the ancient MADBUB kinase domain, or acquired new activity as a result of neofunctionalization, that might explain its conservation during vertebrate evolution.…”

Section: Discussionmentioning

confidence: 99%

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The BUBR1 pseudokinase domain promotes efficient kinetochore PP2A-B56 recruitment to regulate spindle checkpoint silencing and chromosome alignment

Braga

Cisneros²,

Mathieu

et al. 2019

Preprint

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The balance of phospho-signalling at outer-kinetochores during mitosis is critical for the accurate attachments between kinetochores and the mitotic spindle and timely exit from mitosis. In humans, a major player in determining this balance is the PP2A-B56 phosphatase which is recruited to the Kinase Attachment Regulatory Domain (KARD) of the Spindle Assembly Checkpoint protein Budding Uninhibited by Benzimidazole 1-related 1 (BUBR1) in a phospho-dependent manner. This event unleashes a rapid, switch-like phosphatase relay that reverses phosphorylation at the kinetochore, extinguishing the checkpoint and promoting anaphase entry. Here, we conclusively demonstrate that the pseudokinase domain of human BUBR1 lacks phosphotransfer activity and that it was maintained in vertebrates because it allosterically promotes KARD phosphorylation. Mutation or removal of this domain results in decreased PP2A-B56 recruitment to the outer kinetochore, attenuated checkpoint silencing and errors in chromosome alignment as a result of imbalance in Aurora B activity. We demonstrate that the functions of the BUBR1 pseudokinase and the BUB1 kinase domains are intertwined, providing an explanation for retention of the pseudokinase domain in certain eukaryotes.

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Section: Discussionsupporting

confidence: 62%

Section: Resultsmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

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The BUBR1 pseudokinase domain promotes efficient kinetochore PP2A-B56 recruitment to regulate spindle checkpoint silencing and chromosome alignment

Braga

Cisneros²,

Mathieu

et al. 2019

Preprint

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“…Interestingly, this motif has been found to be dispensable for catalytic activity in an atypical catalytically active coccidian kinase, which has been termed WNG1 (with no glycine-1) (43). A recent study suggests a catalytically active, druggable conformation in Drosophila BubR1 that can directly phosphorylate the motor protein centromereassociated protein E (CENP-E) (44). This likely differentiates it from human BubR1, which is reported to be an inactive pseudokinase (45) lacking a Gly-rich loop and that does not bind detectably to ATP (46).…”

Section: Structural Biology As a Key Driver In The Pseudokinase And Pmentioning

confidence: 99%

Emerging concepts in pseudoenzyme classification, evolution, and signaling

et al. 2019

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The 21st century is witnessing an explosive surge in our understanding of pseudoenzyme-driven regulatory mechanisms in biology. Pseudoenzymes are proteins that have sequence homology with enzyme families but that are proven or predicted to lack enzyme activity due to mutations in otherwise conserved catalytic amino acids. The best-studied pseudoenzymes are pseudokinases, although examples from other families are emerging at a rapid rate as experimental approaches catch up with an avalanche of freely available informatics data. Kingdom-wide analysis in prokaryotes, archaea and eukaryotes reveals that between 5 and 10% of proteins that make up enzyme families are pseudoenzymes, with notable expansions and contractions seemingly associated with specific signaling niches. Pseudoenzymes can allosterically activate canonical enzymes, act as scaffolds to control assembly of signaling complexes and their localization, serve as molecular switches, or regulate signaling networks through substrate or enzyme sequestration. Molecular analysis of pseudoenzymes is rapidly advancing knowledge of how they perform noncatalytic functions and is enabling the discovery of unexpected, and previously unappreciated, functions of their intensively studied enzyme counterparts. Notably, upon further examination, some pseudoenzymes have previously unknown enzymatic activities that could not have been predicted a priori. Pseudoenzymes can be targeted and manipulated by small molecules and therefore represent new therapeutic targets (or anti-targets, where intervention should be avoided) in various diseases. In this review, which brings together broad bioinformatics and cell signaling approaches in the field, we highlight a selection of findings relevant to a contemporary understanding of pseudoenzyme-based biology.

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“…10 Settling these discrepancies is particularly important now that BubR1 has been implicated in key biological processes such as maintenance of genomic integrity, cancer, senescence, and aging. In a recent paper published in Cell Research, Huang et al 11 provide compelling evidence that BubR1 is indeed an active kinase. Their first piece of evidence came from resolving the crystal structure of the BubR1 kinase domain.…”

mentioning

confidence: 99%