EVI1 carboxy-terminal phosphorylation is ATM-mediated and sustains transcriptional modulation and self-renewal via enhanced CtBP1 association

Paredes, Roberto; Schneider, Marion; Stevens, Adam; White, Daniel J.; Williamson, Andrew J.K.; Muter, Joanne; Pearson, Stella; Kelly, James R.; Connors, Kathleen; Chadwick, John; Löffler, Harald; Teng, Hsiang Ying; Lovell, Simon C.; Unwin, Richard D.; Vrugt, Henri J. van de; Smith, Hannah; Kustikova, Olga; Schambach, Axel; Somervaille, Tim C. P.; Pierce, Andrew; Whetton, Anthony D.; Meyer, Stefan

doi:10.1093/nar/gky536

Cited by 12 publications

(25 citation statements)

References 51 publications

(72 reference statements)

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“…1b), and in a phosphorylated form at serine S436 (Fig. 1c), in addition to the previously reported EVI1 phosphorylation sites S196, S858 and S860 15,16 . This confirmed EVI1-S436 phosphorylation listed in other studies of cell lines and clinical samples 13,36 .…”

Section: Dynamic Evi1 Phosphorylation At Serine 436 (S436)supporting

confidence: 86%

“…Expression patterns mediated by EVI1-WT include genes with an essential role in stem cell maintenance, and show that EVI1 with S436 available for phosphorylation has a much more coordinated effect on the entire transcriptome, implying that the S436 phosphorylation focuses transcriptional patterns towards self-renewal. Recent data from our group and others suggest that many EVI1 functions are regulated by dynamic interactions with other proteins 6,13,15 . To explain the differences in gene expression patterns mediated by EVI1-WT compared with EVI1-S436 we provide evidence for modulation of EVI1-protein interactions by S436 phosphorylation, with increased affinity of non-phosphorylatable EVI1-S436A to CtBP1.…”

Section: Discussionmentioning

confidence: 90%

“…Substitution of S436 with alanine (A) to create the vector pCMV-EVI1-S436A-flag was done by site-directed mutagenesis using the QuikChange ® II XL Kit (Agilent, Cheadle, UK). The lentiviral vector expressing codon-optimized mouse Evi1-pRRL.PPT.SF.EVI1mCo.IRES_EGFP.pre was mutated as above to generate pRRL.PPT.SF.EVI1mCoS436.IRE-S_EGFP.pre 15,[17][18][19] . Control pRRL.PPT.SF.IRES_EGFP.…”

Section: Plasmids and Site-directed Mutagenesismentioning

confidence: 99%

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EVI1 phosphorylation at S436 regulates interactions with CtBP1 and DNMT3A and promotes self-renewal

et al. 2020

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The transcriptional regulator EVI1 has an essential role in early development and haematopoiesis. However, acute myeloid leukaemia (AML) driven by aberrantly high EVI1 expression has very poor prognosis. To investigate the effects of post-translational modifications on EVI1 function, we carried out a mass spectrometry (MS) analysis of EVI1 in AML and detected dynamic phosphorylation at serine 436 (S436). Wild-type EVI1 (EVI1-WT) with S436 available for phosphorylation, but not non-phosphorylatable EVI1-S436A, conferred haematopoietic progenitor cell self-renewal and was associated with significantly higher organised transcriptional patterns. In silico modelling of EVI1-S436 phosphorylation showed reduced affinity to CtBP1, and CtBP1 showed reduced interaction with EVI1-WT compared with EVI1-S436A. The motif harbouring S436 is a target of CDK2 and CDK3 kinases, which interacted with EVI1-WT. The methyltransferase DNMT3A bound preferentially to EVI1-WT compared with EVI1-S436A, and a hypomethylated cell population associated by EVI1-WT expression in murine haematopoietic progenitors is not maintained with EVI1-S436A. These data point to EVI1-S436 phosphorylation directing functional protein interactions for haematopoietic self-renewal. Targeting EVI1-S436 phosphorylation may be of therapeutic benefit when treating EVI1-driven leukaemia.

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Section: Dynamic Evi1 Phosphorylation At Serine 436 (S436)supporting

confidence: 86%

Section: Discussionmentioning

confidence: 90%

Section: Plasmids and Site-directed Mutagenesismentioning

confidence: 99%

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EVI1 phosphorylation at S436 regulates interactions with CtBP1 and DNMT3A and promotes self-renewal

et al. 2020

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“…The oncogenic function of EVI1 is mediated by its established role on epigenetic regulation and transcriptional control [16][17][18]. EVI1 interacts with Polycomb-group (PcG) proteins to repress the expression of the tumor suppressor gene PTEN [19].…”

Section: Introductionmentioning

confidence: 99%

EVI1 as a Prognostic and Predictive Biomarker of Clear Cell Renal Cell Carcinoma

Palomero

Bodnar

Mateo

et al. 2020

Cancers

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The transcription factor EVI1 plays an oncogenic role in several types of neoplasms by promoting aggressive cancer features. EVI1 contributes to epigenetic regulation and transcriptional control, and its overexpression has been associated with enhanced PI3K-AKT-mTOR signaling in some settings. These observations raise the possibility that EVI1 influences the prognosis and everolimus-based therapy outcome of clear cell renal cell carcinoma (ccRCC). Here, gene expression and protein immunohistochemical studies of ccRCC show that EVI1 overexpression is associated with advanced disease features and with poorer outcome—particularly in the CC-e.3 subtype defined by The Cancer Genome Atlas. Overexpression of an oncogenic EVI1 isoform in RCC cell lines confers substantial resistance to everolimus. The EVI1 rs1344555 genetic variant is associated with poorer survival and greater progression of metastatic ccRCC patients treated with everolimus. This study leads us to propose that evaluation of EVI1 protein or gene expression, and of EVI1 genetic variants may help improve estimates of prognosis and the benefit of everolimus-based therapy in ccRCC.

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“…Lately, CtBPs have also been implied as transcriptional co-repressors which exert diverse functions with regard to tumorigenesis processes [4]. CtBPs, including CtBP1 and CtBP2, were reported to be highly expressed throughout development and revealed as vital supervisors of tissue morphogenesis and organogenesis [5,6]. As an example, CtBP2-null mice exerts a embryonic lethality and often present heart flaws, and insufficient neural development [7,8].…”

Section: Introductionmentioning

confidence: 99%

The C-terminal of the E1A binding protein 2 enhances the metastatic potential of keratinocyte-derived cutaneous squamous carcinoma cells via the gp130/JAK2/Stat1 signaling pathway

Zuo

Bao

et al. 2020

Preprint

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Background The abnormal expression of C-terminal of the E1A binding proteins (CtBPs) was is observed to be involved in several human tumors. This study aims to investigate the expression of CtBPs and their potential underlying mechanisms in the most common metastatic skin cancer, keratinocyte-derived cutaneous squamous cell carcinoma (cSCCs) patients and their evaluation value in clinical diagnosis and prognosis. Methods In the present study, the expression levels of CtBPs in cSCCs and actinic keratosis tissues were examined by immunohistochemical assay. To discover the effects of CtBP2 on the metastatic phenotype of cSCCs, a stably expressing plasmid was transfected into keratinocyte cells in order to initiate CtBP2 overexpression. The endogenous protein gp130, which is a signal-transducing subunit associated with the ligand-occupied interleukin receptor was silenced in these cells using short hairpin RNA (shRNA) sequences. Moreover, the endogenous expression of CtBP2 in cSCCs cell lines was also silenced. Transfection efficiency was validated by western blotting and immunofluorescence assays, and the malignant phenotype of these cells was evaluated by various assays including the CCK8, colony formation, transwell, and cell scratch assay. Furthermore, the activation state of the gp130/JAK2/Stat1 signaling pathway was further discovered via western blotting. Results The observation discovered that the mean levels of CtBP2 expression, not noted for CtBP1, were elevated in 104 cSCCs tissue samples compared with those noted in 102 actinic keratosis tissue samples. The upregulated expression levels of CtBP2 were remarkably associated with tumor metastasis. CtBP2 promoted the malignant phenotype of keratinocyte cells, and gp130 knockdown (k/o) notably reduced this effect in keratinocyte cells by inhibiting the activation of the JAK2/Stat1 pathway. In addition, the k/o of CtBP2 notably reduced the ability of cSCCs cells to metastasize by suppressing the gp130/JAK2/Stat1 pathway. Conclusions The present data revealed that CtBP2 promoted cSCCs cell metastasis via the gp130/JAK2/Stat1 pathway, suggesting that targeting of CtBP2 or gp130 is a promising anti-tumor tactics to impede tumor progression in cSCCs.

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EVI1 carboxy-terminal phosphorylation is ATM-mediated and sustains transcriptional modulation and self-renewal via enhanced CtBP1 association

Cited by 12 publications

References 51 publications

EVI1 phosphorylation at S436 regulates interactions with CtBP1 and DNMT3A and promotes self-renewal

EVI1 phosphorylation at S436 regulates interactions with CtBP1 and DNMT3A and promotes self-renewal

EVI1 as a Prognostic and Predictive Biomarker of Clear Cell Renal Cell Carcinoma

The C-terminal of the E1A binding protein 2 enhances the metastatic potential of keratinocyte-derived cutaneous squamous carcinoma cells via the gp130/JAK2/Stat1 signaling pathway

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