The Canonical Wnt Signal Restricts the Glycogen Synthase Kinase 3/Fbw7-Dependent Ubiquitination and Degradation of Eya1 Phosphatase

Sun, Ye; Li, Xue

doi:10.1128/mcb.00104-14

Cited by 10 publications

(7 citation statements)

References 41 publications

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“…When lysates of cells expressing cMyc or EYA1 alone, or both, were immunoprecipitated using antiFlag-EYA1, cMyc and endogenous FBW7 were also coprecipitated (Fig. 10B), suggesting that FBW7 and EYA1 form a complex in vivo, consistent with the report that FBW7 binds to EYA1 in vivo (24). In sum, these results suggest that EYA1 regulates cMyc stability by affecting the degradation machinery through binding to cMyc and regulation of the phospho-T58-Myc levels, which in turn impairs the recruitment of FBW7 to cMyc, subsequently leading to reduced ubiquitination of Myc.…”

Section: Fig 4 Nmr Analysis (A)supporting

confidence: 84%

EYA1's Conformation Specificity in Dephosphorylating Phosphothreonine in Myc and Its Activity on Myc Stabilization in Breast Cancer

Rodrı́guez

Cheng

et al. 2017

Molecular and Cellular Biology

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EYA1 is known to be overexpressed in human breast cancer, in which the Myc protein is also accumulated in association with decreased phospho-T58 (pT58) levels. We have recently reported that EYA1 functions as a unique protein phosphatase to dephosphorylate Myc at pT58 to regulate Myc levels. However, it remains unclear whether EYA1-mediated Myc dephosphorylation on T58 is a critical function in regulating Myc protein stability in breast cancer. Furthermore, EYA1's substrate specificity has remained elusive. In this study, we have investigated these questions, and here, we report that depletion of EYA1 using short hairpin RNA (shRNA) in breast cancer cells destabilizes the Myc protein and increases pT58 levels, leading to an increase in the doubling time and impairment of cell cycle progression. In correlation with EYA1-mediated stabilization of cMyc and reduced levels of pT58, EYA1 greatly reduced cMyc-FBW7 binding and cMyc ubiquitination, thus providing novel insight into how EYA1 acts to regulate the FBW7-mediated Myc degradation machinery. We found that the conserved C-terminal haloacid dehalogenase domain of EYA1, which has been reported to have only tyrosine phosphatase activity, has dual phosphatase activities, and both the N-and C-terminal domains interact with substrates to increase the catalytic activity of EYA1. Enzymatic assay and nuclear magnetic resonance (NMR) analysis demonstrated that EYA1 has a striking conformation preference for phospho-T58 of Myc. Together, our results not only provide novel structural evidence about the conformation specificity of EYA1 in dephosphorylating phosphothreonine in Myc but also reveal an important mechanism contributing to Myc deregulation in human breast cancer. KEYWORDS EYA1, threonine phosphatase, Myc, degradation, FBW7, cell proliferation, breast cancer, deregulation T he transcription factor Eyes absent (EYA) represents the prototype of a novel class of eukaryotic aspartyl protein tyrosine phosphatases (1-3) that play important roles in development and disease (4-6). While the conserved C-terminal haloacid dehalogenase domain of EYA (ED), which contains the signature motif for the aspartate-based serine/threonine (Ser/Thr) phosphatase family (1-3, 7), has been reported to have only tyrosine phosphatase activity (8), a recent study reported that the N terminus of EYA (NT), which bears no sequence similarity to any known Ser/Thr phosphatase families, has Thr phosphatase activity (8). Therefore, it remains unclear which domain of EYA contains residues for substrate binding and catalytic activity and how EYA achieves

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Section: Fig 4 Nmr Analysis (A)supporting

confidence: 84%

EYA1's Conformation Specificity in Dephosphorylating Phosphothreonine in Myc and Its Activity on Myc Stabilization in Breast Cancer

Rodrı́guez

Cheng

et al. 2017

Molecular and Cellular Biology

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“…EYA activity could be regulated by multiple signaling pathways. Dr. Li and her coworkers demonstrated that hyperactivation of the canonical Wnt and PI3K/Akt signaling pathways reduces EYA1 ubiquitination and thus prevents its premature degradation [85]. In particular, PI3K/Akt signaling was shown to repress the SUMOylation of EYA1 in a phosphorylation-dependent manner and thus enhance the transcriptional activity of EYA1 [86].…”

Section: Potential Strategy Of Targeting the Rdgn Pathway For Clinicamentioning

confidence: 99%

The retinal determination gene network: from developmental regulator to cancer therapeutic target

Kong

Liu

et al. 2016

Oncotarget

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Although originally identified for its function in Drosophila melanogaster eye specification, the Retinal Determination Gene Network (RDGN) is essential for the development of multiple organs in mammals. The RDGN regulates proliferation, differentiation and autocrine signaling, and interacts with other key signaling pathways. Aberrant expression of RDGN members such as DACH, EYA and SIX contributes to tumor initiation and progression; indeed, the levels of RDGN members are clinically prognostic factors in various cancer types. Stimulation or suppression of the activities of these crucial components can block cancer cell proliferation, prevent cancer stem cell expansion and even reverse the EMT process, thereby attenuating malignant phenotypes. Thus, cancer therapeutic interventions targeting RDGN members should be pursued in future studies.

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“…The striking agreement between the peptides and proteins observed in the plasma of breast cancer patients and the previous literature on breast cancer tumors, adjacent fluids, cell lines or blood fluids indicates that LC–ESI–MS/MS of blood peptides will be a powerful tool for selecting plasma proteins and peptides for further research and confirmation. The results of mass spectrometry show striking agreement with previous genetic or biochemical experiments on cancer tissues, tumors, biopsies or cell lines: CPEB1 [63], LTBP4 [64], HIF1A [65, 66], IGHE [67], RAB44 [68], NEFM [39], C19orf82, SLC35B1 [69], 1D12A that shows a cyptic alignment with cyclin-dependent kinase-like isoform 1 [70], C8orf34 [71], OCLN [72], EYA1 [73], HLA-DRB1 [74], LAR [75] and LRRC4B that interacts with the LARS receptor phosphatases [76], PTPDC1 [77], WWC1 [78], ZNF562, PTMA [79], MGAT1 [80], NDUFA1 [81], NOGOC [82], olfactory receptors OR1E or the HSA12 protein [83], GCSH [84], ELTD1 [85], TBX15 [86], orphan nuclear receptors such as NR2C2 [87], autophagy related proteins such as ATG16L1 (FLJ00045) that regulate the production of extracellular vesicles called exosomes [88], PDLIM1 [89, 90], GALNT9 [91], ASH2L [92], PPFIBP1 [93], SLCO3A1 [94], BHMT2 [95], CS citrate synthase [96] FAM188B2 inactive ubiquitin carboxyl-terminal hydrolase MINDY4B that is expressed in breast cancer tissue, LGALS7 [97] SAT2 [98], SFRS8, SLC22A12 [99], WNT9B [100], SLC2A4 [101], ZNF101, WT1 (Wilms Tumor Protein) [102], CCDC47 [103], ERLIN1 (SPFH1) and MREG [104], EID2 [105], THOC1 [106, 107], DDX47 [108], PTPRE [109], EMILIN1 [110], DKFZp779G1236 (piccolo, or piBRCA2) [111], MAP3K8 [112] regulated by Serine/Arginine-Rich Splicing Factor Kinase [113], QSER1 [39], IQCJ-SCHIP1 [114, 115], ANXA4 [116] and DHDDS [117] among others. The disease-specific proteins and peptides may result from the introduction of new proteins into cir...…”

Section: Discussionmentioning

confidence: 99%

The plasma peptides of breast versus ovarian cancer

et al. 2019

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BackgroundThere is a need to demonstrate a proof of principle that proteomics has the capacity to analyze plasma from breast cancer versus other diseases and controls in a multisite clinical trial design. The peptides or proteins that show a high observation frequency, and/or precursor intensity, specific to breast cancer plasma might be discovered by comparison to other diseases and matched controls. The endogenous tryptic peptides of breast cancer plasma were compared to ovarian cancer, female normal, sepsis, heart attack, Alzheimer’s and multiple sclerosis along with the institution-matched normal and control samples collected directly onto ice.MethodsEndogenous tryptic peptides were extracted from individual breast cancer and control EDTA plasma samples in a step gradient of acetonitrile, and collected over preparative C18 for LC–ESI–MS/MS with a set of LTQ XL linear quadrupole ion traps working together in parallel to randomly and independently sample clinical populations. The MS/MS spectra were fit to fully tryptic peptides or phosphopeptides within proteins using the X!TANDEM algorithm. The protein observation frequency was counted using the SEQUEST algorithm after selecting the single best charge state and peptide sequence for each MS/MS spectra. The observation frequency was subsequently tested by Chi Square analysis. The log10 precursor intensity was compared by ANOVA in the R statistical system.ResultsPeptides and/or phosphopeptides of common plasma proteins such as APOE, C4A, C4B, C3, APOA1, APOC2, APOC4, ITIH3 and ITIH4 showed increased observation frequency and/or precursor intensity in breast cancer. Many cellular proteins also showed large changes in frequency by Chi Square (χ2 > 100, p < 0.0001) in the breast cancer samples such as CPEB1, LTBP4, HIF-1A, IGHE, RAB44, NEFM, C19orf82, SLC35B1, 1D12A, C8orf34, HIF1A, OCLN, EYA1, HLA-DRB1, LARS, PTPDC1, WWC1, ZNF562, PTMA, MGAT1, NDUFA1, NOGOC, OR1E1, OR1E2, CFI, HSA12, GCSH, ELTD1, TBX15, NR2C2, FLJ00045, PDLIM1, GALNT9, ASH2L, PPFIBP1, LRRC4B, SLCO3A1, BHMT2, CS, FAM188B2, LGALS7, SAT2, SFRS8, SLC22A12, WNT9B, SLC2A4, ZNF101, WT1, CCDC47, ERLIN1, SPFH1, EID2, THOC1, DDX47, MREG, PTPRE, EMILIN1, DKFZp779G1236 and MAP3K8 among others. The protein gene symbols with large Chi Square values were significantly enriched in proteins that showed a complex set of previously established functional and structural relationships by STRING analysis. An increase in mean precursor intensity of peptides was observed for QSER1 as well as SLC35B1, IQCJ-SCHIP1, MREG, BHMT2, LGALS7, THOC1, ANXA4, DHDDS, SAT2, PTMA and FYCO1 among others. In contrast, the QSER1 peptide QPKVKAEPPPK was apparently specific to ovarian cancer.ConclusionThere was striking agreement between the breast cancer plasma peptides and proteins discovered by LC–ESI–MS/MS with previous biomarkers from tumors, cells lines or body fluids by genetic or biochemical methods. The results indicate that variation in plasma peptides from breast cancer versus ovarian cancer may be directly discovered by LC–ESI–MS/...

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The Canonical Wnt Signal Restricts the Glycogen Synthase Kinase 3/Fbw7-Dependent Ubiquitination and Degradation of Eya1 Phosphatase

Cited by 10 publications

References 41 publications

EYA1's Conformation Specificity in Dephosphorylating Phosphothreonine in Myc and Its Activity on Myc Stabilization in Breast Cancer

EYA1's Conformation Specificity in Dephosphorylating Phosphothreonine in Myc and Its Activity on Myc Stabilization in Breast Cancer

The retinal determination gene network: from developmental regulator to cancer therapeutic target

The plasma peptides of breast versus ovarian cancer

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