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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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Autores principales: Belsham, Hannah R., Friel, Claire T.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5723132/
https://www.ncbi.nlm.nih.gov/pubmed/29230353
http://dx.doi.org/10.7717/peerj.4034
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author Belsham, Hannah R.
Friel, Claire T.
author_facet Belsham, Hannah R.
Friel, Claire T.
author_sort Belsham, Hannah R.
collection PubMed
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.
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spelling pubmed-57231322017-12-11 A Cdk1 phosphomimic mutant of MCAK impairs microtubule end recognition Belsham, Hannah R. Friel, Claire T. PeerJ Biochemistry 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 Inc. 2017-12-06 /pmc/articles/PMC5723132/ /pubmed/29230353 http://dx.doi.org/10.7717/peerj.4034 Text en ©2017 Belsham and Friel http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Biochemistry
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 Biochemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5723132/
https://www.ncbi.nlm.nih.gov/pubmed/29230353
http://dx.doi.org/10.7717/peerj.4034
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