Microtubule regulation in mitosis: tubulin phosphorylation by the cyclin-dependent kinase Cdk1.

Abstract:

:The activation of the cyclin-dependent kinase Cdk1 at the transition from interphase to mitosis induces important changes in microtubule dynamics. Cdk1 phosphorylates a number of microtubule- or tubulin-binding proteins but, hitherto, tubulin itself has not been detected as a Cdk1 substrate. Here we show that Cdk1 phosphorylates beta-tubulin both in vitro and in vivo. Phosphorylation occurs on Ser172 of beta-tubulin, a site that is well conserved in evolution. Using a phosphopeptide antibody, we find that a fraction of the cell tubulin is phosphorylated during mitosis, and this tubulin phosphorylation is inhibited by the Cdk1 inhibitor roscovitine. In mitotic cells, phosphorylated tubulin is excluded from microtubules, being present in the soluble tubulin fraction. Consistent with this distribution in cells, the incorporation of Cdk1-phosphorylated tubulin into growing microtubules is impaired in vitro. Additionally, EGFP-beta3-tubulin(S172D/E) mutants that mimic phosphorylated tubulin are unable to incorporate into microtubules when expressed in cells. Modeling shows that the presence of a phosphoserine at position 172 may impair both GTP binding to beta-tubulin and interactions between tubulin dimers. These data indicate that phosphorylation of tubulin by Cdk1 could be involved in the regulation of microtubule dynamics during mitosis.

journal_name

Mol Biol Cell

authors

Fourest-Lieuvin A,Peris L,Gache V,Garcia-Saez I,Juillan-Binard C,Lantez V,Job D

doi

10.1091/mbc.e05-07-0621

keywords:

subject

Has Abstract

pub_date

2006-03-01 00:00:00

pages

1041-50

issue

3

eissn

1059-1524

issn

1939-4586

pii

E05-07-0621

journal_volume

17

pub_type

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