Dataset Information

Greatwall kinase regulates SAC by controlling phosphorylation and activity of MPS1

ABSTRACT: The Greatwall/Mastl kinase is an essential gene, which regulates the phosphatase activity that counteracts the phosphorylation of Cdk1 substrates. Although Mastl-/- MEFs can enter mitosis, they are unable to complete mitosis due to chromosome segregation defects. Here, we show that Mastl is essential for robust spindle assembly checkpoint (SAC) maintenance. Mastl-/- MEFs display reduced phosphorylation and kinase activity of MPS1, which causes mislocalization of MPS1, Mad1, and Aurora B from the kinetochore and the inner centromere regions. This results in premature inactivation of SAC signalling and early anaphase onset without correction of chromosome-spindle attachment mistakes. Treatment of the knockout cells with protein phosphatase II inhibitor okadaic acid (OA) rescued decreased MPS1 activity, mislocalization of Aurora B, Mad1, MPS1, and premature mitotic slippage. Our data demonstrates how regulation of PP2A activity by Mastl kinase prevents premature SAC silencing as a result of controlling the phosphorylation status and activity of MPS1.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Sheena Wee

LAB HEAD: Jayantha Gunaratne

PROVIDER: PXD004882 | Pride | 2018-10-26

REPOSITORIES: Pride

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Publications

Loss of the Greatwall Kinase Weakens the Spindle Assembly Checkpoint.

Diril M Kasim MK Bisteau Xavier X Kitagawa Mayumi M Caldez Matias J MJ Wee Sheena S Gunaratne Jayantha J Lee Sang Hyun SH Kaldis Philipp P

PLoS genetics 20160915 9

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 ...[more]

PMID: 27631493

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Project description:MASTL (Microtubule associated serine-threonine kinase-like) is a relatively recently identified mitotic accelerator. There are also indications of its role in cancer progression, especially in breast cancer, but the molecular mechanisms behind this and the cell cycle independent functions of MASTL are poorly understood. Regulation of cell adhesion and actin dynamics play a central role in governing cell contractility, migration and cell division in normal and cancer cells. Here, we identify MASTL as regulator of cell contractility and activator of the transcriptional co-activator MTRF-A. We show that depletion of MASTL increases cell contact with the extracellular matrix, reduces contractile actin stress fibers in normal and breast cancer cells. Importantly, depletion of MASTL also leads to strongly impairs motility of breast cancer cells. Our transcriptome and proteome profiling revealed that depletion of MASTL impairs transcription and expression of several MRTF-A target genes implicated in cellular movement and actomyosin contraction, including Rho Guanine nucleotide exchange factor 2 (GEF-H1, ARHGEF2) and tropomyosin 4.2 (TPM4). Mechanistically, we find that MASTL is necessary for serum-induced activation of SRF/MTRF-A transcription and that exogenous expression of GEF-H1 in MASTL depleted cells is sufficient to restore cell contractility. Taken together, our results suggest that MASTL is a previously undescribed key regulator of cell morphology and the actin cytoskeleton through transcriptional control of multiple adhesion and actin cytoskeleton regulating genes with key roles in contractility, cell adhesion and migration. MASTL (Microtubule associated serine-threonine kinase-like) is a relatively recently identified mitotic accelerator. There are also indications of its role in cancer progression, especially in breast cancer, but the molecular mechanisms behind this and the cell cycle independent functions of MASTL are poorly understood. Regulation of cell adhesion and actin dynamics play a central role in cell migration, morphology and cancer progression, but the role of MASTL in these processes has not been evaluated. Here, we show that depletion of MASTL increases cell contact with the extracellular matrix in breast cancer cells. Importantly, depletion of MASTL also leads to reduction of contractile actin stress fibers and decreased cell motility. Further, our transcriptome and proteome profiling revealed that mechanistically depletion of MASTL impairs transcription and expression of regulators of cellular movement and actomyosin contraction, including Rho Guanine nucleotide exchange factor 2 (GEF-H1, ARHGEF2) and tropomyosin 4.2 (TPM4). Further, we show that the transcriptional changes are caused by defectiveness of the serum response factor (SRF) signalling. Importantly, this study reveals advanced role for MASTL in interphase cells that activates a transcriptional program strongly impacting on the expression of regulators of cellular motility and actomyosin contraction.

Dataset Information

Greatwall kinase regulates SAC by controlling phosphorylation and activity of MPS1

Publications

Loss of the Greatwall Kinase Weakens the Spindle Assembly Checkpoint.

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