Dataset Information

Microtubule-associated serine/threonine-protein kinase-like (MASTL) surfaceome proteomics analysis

ABSTRACT: Microtubule-associated serine/threonine-protein kinase-like (MASTL; a.k.a. Greatwall (Gwl) kinase) has an established role in the acceleration of cell cycle progression through inhibition of the PP2A-B55 phosphatase activity. Importantly, MASTL has emerged as a putative oncogene. Especially in breast cancer, high MASTL expression levels correlate with increased tumor growth and metastasis in vivo. We reported recently a kinase-independent role for MASTL as a regulator of cell adhesion, contractility and MRTF-A/SRF activity in breast cancer. This study also revealed that depletion of MASTL affects transcription and protein expression of multiple targets. Cell surface proteins are critical for characterization, isolation, and regulation of signaling. In order to reveal if MASTL affects breast cancer cells directly by regulation of surface proteins, we performed SILAC based surfaceome proteomics. Heavy and light isotope labelled endogenously produced surface proteins of MDA-MB-231 cells were purified by biotin enrichment and subjected to proteomics analysis.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Breast

DISEASE(S): Breast Cancer

SUBMITTER: Sergio Lilla

LAB HEAD: Sara Rossana Zanivan

PROVIDER: PXD019993 | Pride | 2022-06-28

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	20180420_SEL_HF_SA_ELN_R20_100418_MASTL-H-NT-L_fwd_C_001.raw	Raw
	20180420_SEL_HF_SA_ELN_R20_100418_MASTL-H-NT-L_fwd_C_002.raw	Raw
	20180420_SEL_HF_SA_ELN_R20_100418_MASTL-H-NT-L_fwd_C_003.raw	Raw
	20180420_SEL_HF_SA_ELN_R20_100418_MASTL-H-NT-L_fwd_C_004.raw	Raw
	20180420_SEL_HF_SA_ELN_R20_100418_MASTL-H-NT-L_fwd_C_005.raw	Raw

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Publications

MASTL is enriched in cancerous and pluripotent stem cells and influences OCT1/OCT4 levels.

Närvä Elisa E Taskinen Maria E ME Lilla Sergio S Isomursu Aleksi A Pietilä Mika M Weltner Jere J Isola Jorma J Sihto Harri H Joensuu Heikki H Zanivan Sara S Norman Jim J Ivaska Johanna J

iScience 20220525 6

MASTL is a mitotic accelerator with an emerging role in breast cancer progression. However, the mechanisms behind its oncogenicity remain largely unknown. Here, we identify a previously unknown role and eminent expression of MASTL in stem cells. MASTL staining from a large breast cancer patient cohort indicated a significant association with β3 integrin, an established mediator of breast cancer stemness. MASTL silencing reduced OCT4 levels in human pluripotent stem cells and OCT1 in breast cance ...[more]

PMID: 35677646

Similar Datasets

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.

Project description:Breast cancer is a heterogeneous disease comprised of at least five major subtypes. Luminal subtype tumors confer a more favourable patient prognosis, which is in part, attributed to the Estrogen Receptor-alpha (ER) positivity and anti-hormone responsiveness of these tumors. Expression of the forkhead box transcription factor, FOXA1, also correlates with the luminal subtype and patient survival, but is present in a subset of ER-negative tumors. Similarly, FOXA1 is consistently expressed in luminal breast cancer cell lines even in the absence of ER. In contrast, basal breast cancer cell lines do not express FOXA1, and loss of FOXA1 in luminal cells increases migration and invasion, characteristics of the basal subtype. To delineate an ER-independent role for FOXA1 in maintaining the luminal phenotype, and hence a more favourable prognosis, we performed cDNA microarray analyses on luminal FOXA1-positive, ER-positive (MCF7, T47D) and FOXA1-positive, ER-negative (MDA-MB-453, SKBR3) cell lines in the presence or absence of transient FOXA1 silencing. This resulted in three FOXA1 transcriptomes: (1) a luminal-signature (consistent across cell lines), (2) an ER-positive signature (restricted to MCF7 and T47D) and (3) an ER-negative signature (restricted to MDA-MB-453 and SKBR3). Use of Gene Set Enrichment Analyses (GSEA) as a phenotyping tool revealed that FOXA1 silencing resulted in a transcriptome shift from luminal to basal gene expression signatures. FOXA1 binds to both luminal and basal genes within luminal breast cancer cells, suggesting that it not only transactivates luminal genes, but also represses basal-associated genes. From these results we conclude that FOXA1 controls plasticity between basal and luminal cells, playing a dominant role in repressing the basal phenotype, and thus tumor aggressiveness, in luminal breast cancer cells. Although it has been proposed that FOXA1-targeting agents may be useful for treating luminal tumors, these data suggest that this approach may promote transitions toward a more aggressive cancer. FOXA1 siRNA treated breast cell lines compared directly to nonspecific siRNA treated cell lines using Agilent 4X44 microarrays.