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 me...

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Bibliographic Details
Published in:PeerJ (San Francisco, CA) CA), 2017-12, Vol.5, p.e4034-e4034, Article e4034
Main Authors: Belsham, Hannah R, Friel, Claire T
Format: Article
Language:English
Subjects:
Analysis
Biochemistry
Biophysics
Cell cycle
Chromosomes
Cytological research
Depolymerisation
Kinases
Kinesin
Kinesin-13
Localization
MCAK
Metaphase
Microtubule
Microtubule end recognition
Microtubules
Molecular Biology
Mutation
Phosphomimic
Phosphorylation
Properties
Standard deviation
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Summary: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.
ISSN:2167-8359
2167-8359
DOI:10.7717/peerj.4034