Loss of the Greatwall Kinase Weakens the Spindle Assembly Checkpoint

Research output: Contribution to journalArticle

Abstract

The Greatwall kinase/Mastl is an essential gene that indirectly inhibits the phosphatase activity toward mitotic Cdk1 substrates. Here we show that although Mastl knockout (MastlNULL) MEFs enter mitosis, they progress through mitosis without completing cytokinesis despite the presence of misaligned chromosomes, which causes chromosome segregation defects. Furthermore, we uncover the requirement of Mastl for robust spindle assembly checkpoint (SAC) maintenance since the duration of mitotic arrest caused by microtubule poisons in MastlNULL MEFs is shortened, which correlates with premature disappearance of the essential SAC protein Mad1 at the kinetochores. Notably, MastlNULL MEFs display reduced phosphorylation of a number of proteins in mitosis, which include the essential SAC kinase MPS1. We further demonstrate that Mastl is required for multi-site phosphorylation of MPS1 as well as robust MPS1 kinase activity in mitosis. In contrast, treatment of MastlNULL cells with the phosphatase inhibitor okadaic acid (OKA) rescues the defects in MPS1 kinase activity, mislocalization of phospho-MPS1 as well as Mad1 at the kinetochore, and premature SAC silencing. Moreover, using in vitro dephosphorylation assays, we demonstrate that Mastl promotes persistent MPS1 phosphorylation by inhibiting PP2A/B55-mediated MPS1 dephosphorylation rather than affecting Cdk1 kinase activity. Our findings establish a key regulatory function of the Greatwall kinase/Mastl->PP2A/B55 pathway in preventing premature SAC silencing.

Details

Authors
  • M. Kasim Diril
  • Xavier Bisteau
  • Mayumi Kitagawa
  • Matias J. Caldez
  • Sheena Wee
  • Jayantha Gunaratne
  • Sang Hyun Lee
  • Philipp Kaldis
External organisations
  • A*Star Institute of Molecular and Cell Biology (IMCB)
  • Dokuz Eylül University
  • Duke–NUS Medical School
  • National University of Singapore
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Medical Genetics
Original languageEnglish
Article numbere1006310
Pages (from-to)1-26
JournalPLoS Genetics
Volume12
Issue number9
Publication statusPublished - 2016 Sep 15
Publication categoryResearch
Peer-reviewedYes
Externally publishedYes