A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition
The microtubule depolymerising kinesin-13, MCAK, is phosphorylated at residue T537 by Cdk1. This is the only known phosphorylation site within MCAK’s motor domain. To understand the impact of phosphorylation by Cdk1 on microtubule depolymerisation activity, we have investigated the molecular mechani...
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| Format: | Article |
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PeerJ
2017
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| Online Access: | https://eprints.nottingham.ac.uk/48216/ |
| _version_ | 1848797716713308160 |
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| author | Belsham, Hannah R. Friel, Claire T. |
| author_facet | Belsham, Hannah R. Friel, Claire T. |
| author_sort | Belsham, Hannah R. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | The microtubule depolymerising kinesin-13, MCAK, is phosphorylated at residue T537 by Cdk1. This is the only known phosphorylation site within MCAK’s motor domain. To understand the impact of phosphorylation by Cdk1 on microtubule depolymerisation activity, we have investigated the molecular mechanism of the phosphomimic mutant T537E. This mutant significantly impairs microtubule depolymerisation activity and when transfected into cells causes metaphase arrest and misaligned chromosomes. We show that the molecular mechanism underlying the reduced depolymerisation activity of this phosphomimic mutant is an inability to recognise the microtubule end. The microtubule-end residence time is reduced relative to wild-type MCAK, whereas the lattice residence time is unchanged by the phosphomimic mutation. Further, the microtubule-end specific stimulation of ADP dissociation, characteristic of MCAK, is abolished by this mutation. Our data shows that T537E is unable to distinguish between the microtubule end and the microtubule lattice. |
| first_indexed | 2025-11-14T20:08:18Z |
| format | Article |
| id | nottingham-48216 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| last_indexed | 2025-11-14T20:08:18Z |
| publishDate | 2017 |
| publisher | PeerJ |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-482162020-05-04T19:21:16Z https://eprints.nottingham.ac.uk/48216/ A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition Belsham, Hannah R. Friel, Claire T. The microtubule depolymerising kinesin-13, MCAK, is phosphorylated at residue T537 by Cdk1. This is the only known phosphorylation site within MCAK’s motor domain. To understand the impact of phosphorylation by Cdk1 on microtubule depolymerisation activity, we have investigated the molecular mechanism of the phosphomimic mutant T537E. This mutant significantly impairs microtubule depolymerisation activity and when transfected into cells causes metaphase arrest and misaligned chromosomes. We show that the molecular mechanism underlying the reduced depolymerisation activity of this phosphomimic mutant is an inability to recognise the microtubule end. The microtubule-end residence time is reduced relative to wild-type MCAK, whereas the lattice residence time is unchanged by the phosphomimic mutation. Further, the microtubule-end specific stimulation of ADP dissociation, characteristic of MCAK, is abolished by this mutation. Our data shows that T537E is unable to distinguish between the microtubule end and the microtubule lattice. PeerJ 2017-12-06 Article PeerReviewed Belsham, Hannah R. and Friel, Claire T. (2017) A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition. PeerJ, 5 . e4034/1-e4034/12. ISSN 2167-8359 MCAK; kinesin-13; Microtubule; Depolymerisation; Microtubule end recognition; Phosphomimic https://peerj.com/articles/4034/ doi:10.7717/peerj.4034 doi:10.7717/peerj.4034 |
| spellingShingle | MCAK; kinesin-13; Microtubule; Depolymerisation; Microtubule end recognition; Phosphomimic Belsham, Hannah R. Friel, Claire T. A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition |
| title | A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition |
| title_full | A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition |
| title_fullStr | A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition |
| title_full_unstemmed | A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition |
| title_short | A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition |
| title_sort | cdk1 phosphomimic mutant of mcak impairs microtubule end recognition |
| topic | MCAK; kinesin-13; Microtubule; Depolymerisation; Microtubule end recognition; Phosphomimic |
| url | https://eprints.nottingham.ac.uk/48216/ https://eprints.nottingham.ac.uk/48216/ https://eprints.nottingham.ac.uk/48216/ |