Home LiteratureArticle Details
PMID: 30022044 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

p38-mediated phosphorylation at T367 induces EZH2 cytoplasmic localization to promote breast cancer metastasis.

Nature communications ·Vol. 9 ·No. 1 ·2018-00-18 ·Pages 2801

Anwar T, Arellano-Garcia C, Ropa J, Chen YC, Kim HS, Yoon E, Grigsby S, Basrur V, Nesvizhskii AI, Muntean A, Gonzalez ME, Kidwell KM, Nikolovska-Coleska Z, Kleer CG

Abstract

Overexpression of EZH2 in estrogen receptor negative (ER-) breast cancer promotes metastasis. EZH2 has been mainly studied as the catalytic component of the Polycomb Repressive Complex 2 (PRC2) that mediates gene repression by trimethylating histone H3 at lysine 27 (H3K27me3). However, how EZH2 drives metastasis despite the low H3K27me3 levels observed in ER- breast cancer is unknown. Here we show that in human invasive carcinomas and distant metastases, cytoplasmic EZH2 phosphorylated at T367 is significantly associated with ER- disease and low H3K27me3 levels. p38-mediated EZH2 phosphorylation at T367 promotes EZH2 cytoplasmic localization and potentiates EZH2 binding to vinculin and other cytoskeletal regulators of cell migration and invasion. Ectopic expression of a phospho-deficient T367A-EZH2 mutant is sufficient to inhibit EZH2 cytoplasmic expression, disrupt binding to cytoskeletal regulators, and reduce EZH2-mediated adhesion, migration, invasion, and development of spontaneous metastasis. These results point to a PRC2-independent non-canonical mechanism of EZH2 pro-metastatic function.

MeSH Terms
Animals Breast Neoplasms/genetics,mortality,pathology,therapy Carcinoma, Ductal, Breast/genetics,mortality,secondary,therapy Cell Line, Tumor Cell Movement Enhancer of Zeste Homolog 2 Protein/antagonists & inhibitors,genetics,metabolism Estrogen Receptor alpha/genetics,metabolism Female Gene Expression Regulation, Neoplastic Heterografts Histones/genetics,metabolism Humans Lung Neoplasms/genetics,mortality,secondary,therapy Mice Mice, SCID Phosphorylation Polycomb Repressive Complex 2/genetics,metabolism RNA, Small Interfering/genetics,metabolism Survival Analysis Threonine p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors,genetics,metabolism
Chemicals
ESR1 protein, human Estrogen Receptor alpha Histones RNA, Small Interfering Threonine EZH2 protein, human Enhancer of Zeste Homolog 2 Protein Polycomb Repressive Complex 2 p38 Mitogen-Activated Protein Kinases
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
Anwar Talha ORCID
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. | Molecular Cellular and Pathology Training Program, University of Michigan, Ann Arbor, MI, 48109, USA. | Rogel Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA. | Medical Scientist Training Program, University of Michigan, Ann Arbor, MI, 48109, USA.
Arellano-Garcia Caroline
Michigan Post-baccalaureate Research Education Program, University of Michigan, Ann Arbor, MI, 48109, USA.
Ropa James
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. | Molecular Cellular and Pathology Training Program, University of Michigan, Ann Arbor, MI, 48109, USA.
Chen Yu-Chih
Rogel Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA. | Department of Electrical Engineering and Computer Science, University of Michigan, Ann Arbor, MI, 48109, USA.
Kim Hong Sun
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. | Rogel Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA.
Yoon Euisik
Department of Electrical Engineering and Computer Science, University of Michigan, Ann Arbor, MI, 48109, USA. | Department of Biomedical Engineering, University of Michigan, Ann Arbor, MI, 48109, USA.
Grigsby Sierrah
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. | Molecular Cellular and Pathology Training Program, University of Michigan, Ann Arbor, MI, 48109, USA.
Basrur Venkatesha ORCID
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA.
Nesvizhskii Alexey I ORCID
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA.
Muntean Andrew
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA.
Gonzalez Maria E
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. | Rogel Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA.
Kidwell Kelley M
Department of Biostatistics, University of Michigan, Ann Arbor, MI, 48109, USA.
Nikolovska-Coleska Zaneta
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA.
Kleer Celina G
Department of Pathology, University of Michigan Medical School, Ann Arbor, MI, 48109, USA. kleer@umich.edu. | Rogel Cancer Center, University of Michigan, Ann Arbor, MI, 48109, USA. kleer@umich.edu.
References (52)
52 references, click to expand
  1. Small Nucleolar Noncoding RNA SNORA23, Up-Regulated in Human Pancreatic Ductal Adenocarcinoma, Regulates Expression of Spectrin Repeat-Containing Nuclear Envelope 2 to Promote Growth and Metastasis of Xenograft Tumors in Mice.
    Gastroenterology. 2017 Jul;153(1):292-306.e2 PMID: 28390868
  2. Targeting EZH2 in cancer.
    Nat Med. 2016 Feb;22(2):128-34 PMID: 26845405
  3. The methyltransferase Ezh2 controls cell adhesion and migration through direct methylation of the extranuclear regulatory protein talin.
    Nat Immunol. 2015 May;16(5):505-16 PMID: 25751747
  4. TNF/p38α/polycomb signaling to Pax7 locus in satellite cells links inflammation to the epigenetic control of muscle regeneration.
    Cell Stem Cell. 2010 Oct 8;7(4):455-69 PMID: 20887952
  5. Force engages vinculin and promotes tumor progression by enhancing PI3K activation of phosphatidylinositol (3,4,5)-triphosphate.
    Cancer Res. 2014 Sep 1;74(17):4597-611 PMID: 25183785
  6. Scaling and automation of a high-throughput single-cell-derived tumor sphere assay chip.
    Lab Chip. 2016 Oct 7;16(19):3708-17 PMID: 27510097
  7. Inhibition of p38 MAPK sensitizes tumour cells to cisplatin-induced apoptosis mediated by reactive oxygen species and JNK.
    EMBO Mol Med. 2013 Nov;5(11):1759-74 PMID: 24115572
  8. 2016 update of the PRIDE database and its related tools.
    Nucleic Acids Res. 2016 Jan 4;44(D1):D447-56 PMID: 26527722
  9. Invasive breast carcinomas in Ghana: high frequency of high grade, basal-like histology and high EZH2 expression.
    Breast Cancer Res Treat. 2012 Aug;135(1):59-66 PMID: 22527102
  10. SAINTexpress: improvements and additional features in Significance Analysis of INTeractome software.
    J Proteomics. 2014 Apr 4;100:37-43 PMID: 24513533
  11. Step-by-step quantitative analysis of focal adhesions.
    MethodsX. 2014 Jul 07;1:56-9 PMID: 26150935
  12. EZH2 expands breast stem cells through activation of NOTCH1 signaling.
    Proc Natl Acad Sci U S A. 2014 Feb 25;111(8):3098-103 PMID: 24516139
  13. WISP3 (CCN6) is a secreted tumor-suppressor protein that modulates IGF signaling in inflammatory breast cancer.
    Neoplasia. 2004 Mar-Apr;6(2):179-85 PMID: 15140407
  14. Praja1 E3 ubiquitin ligase promotes skeletal myogenesis through degradation of EZH2 upon p38α activation.
    Nat Commun. 2017 Jan 09;8:13956 PMID: 28067271
  15. Invadosomes at a glance.
    J Cell Sci. 2009 Sep 1;122(Pt 17):3009-13 PMID: 19692587
  16. Activation and signaling of the p38 MAP kinase pathway.
    Cell Res. 2005 Jan;15(1):11-8 PMID: 15686620
  17. Combined cDNA array comparative genomic hybridization and serial analysis of gene expression analysis of breast tumor progression.
    Cancer Res. 2006 Apr 15;66(8):4065-78 PMID: 16618726
  18. The CRAPome: a contaminant repository for affinity purification-mass spectrometry data.
    Nat Methods. 2013 Aug;10(8):730-6 PMID: 23921808
  19. GSK3β inactivation promotes the oncogenic functions of EZH2 and enhances methylation of H3K27 in human breast cancers.
    Oncotarget. 2016 Aug 30;7(35):57131-57144 PMID: 27494834
  20. Inhibition of the p38 kinase suppresses the proliferation of human ER-negative breast cancer cells.
    Cancer Res. 2009 Dec 1;69(23):8853-61 PMID: 19920204
  21. Single-cell Migration Chip for Chemotaxis-based Microfluidic Selection of Heterogeneous Cell Populations.
    Sci Rep. 2015 May 18;5:9980 PMID: 25984707
  22. SAINT: probabilistic scoring of affinity purification-mass spectrometry data.
    Nat Methods. 2011 Jan;8(1):70-3 PMID: 21131968
  23. Suppression of tumorigenicity in transformed cells after transfection with vinculin cDNA.
    J Cell Biol. 1992 Oct;119(2):427-38 PMID: 1400584
  24. Characterization of LY2228820 dimesylate, a potent and selective inhibitor of p38 MAPK with antitumor activity.
    Mol Cancer Ther. 2014 Feb;13(2):364-74 PMID: 24356814
  25. Mesenchymal Stem Cell-Induced DDR2 Mediates Stromal-Breast Cancer Interactions and Metastasis Growth.
    Cell Rep. 2017 Jan 31;18(5):1215-1228 PMID: 28147276
  26. Vinculin facilitates cell invasion into three-dimensional collagen matrices.
    J Biol Chem. 2010 Apr 23;285(17):13121-30 PMID: 20181946
  27. Expression of the adherens junction protein vinculin in human basal and squamous cell tumors: relationship to invasiveness and metastatic potential.
    Hum Pathol. 1997 Nov;28(11):1230-6 PMID: 9385927
  28. EZH2 is a marker of aggressive breast cancer and promotes neoplastic transformation of breast epithelial cells.
    Proc Natl Acad Sci U S A. 2003 Sep 30;100(20):11606-11 PMID: 14500907
  29. Somatic mutations altering EZH2 (Tyr641) in follicular and diffuse large B-cell lymphomas of germinal-center origin.
    Nat Genet. 2010 Feb;42(2):181-5 PMID: 20081860
  30. Histone methyltransferase EZH2 induces Akt-dependent genomic instability and BRCA1 inhibition in breast cancer.
    Cancer Res. 2011 Mar 15;71(6):2360-70 PMID: 21406404
  31. Global H3K27 trimethylation and EZH2 abundance in breast tumor subtypes.
    Mol Oncol. 2012 Oct;6(5):494-506 PMID: 22766277
  32. The Polycomb Group protein EZH2 regulates actin polymerization in human prostate cancer cells.
    Prostate. 2008 Feb 15;68(3):255-63 PMID: 18095286
  33. EZH2 oncogenic activity in castration-resistant prostate cancer cells is Polycomb-independent.
    Science. 2012 Dec 14;338(6113):1465-9 PMID: 23239736
  34. Vinculin modulation of paxillin-FAK interactions regulates ERK to control survival and motility.
    J Cell Biol. 2004 May 10;165(3):371-81 PMID: 15138291
  35. The polycomb group protein EZH2 is involved in progression of prostate cancer.
    Nature. 2002 Oct 10;419(6907):624-9 PMID: 12374981
  36. Association of H3K9me3 and H3K27me3 repressive histone marks with breast cancer subtypes in the Nurses' Health Study.
    Breast Cancer Res Treat. 2014 Oct;147(3):639-51 PMID: 25224916
  37. Polycomb group protein ezh2 controls actin polymerization and cell signaling.
    Cell. 2005 May 6;121(3):425-36 PMID: 15882624
  38. Vinculin, cell mechanics and tumour cell invasion.
    Cell Biol Int. 2013 May;37(5):397-405 PMID: 23494917
  39. Phosphorylation of EZH2 activates STAT3 signaling via STAT3 methylation and promotes tumorigenicity of glioblastoma stem-like cells.
    Cancer Cell. 2013 Jun 10;23(6):839-52 PMID: 23684459
  40. An ERK-p38 subnetwork coordinates host cell apoptosis and necrosis during coxsackievirus B3 infection.
    Cell Host Microbe. 2013 Jan 16;13(1):67-76 PMID: 23332156
  41. The eps8 family of proteins links growth factor stimulation to actin reorganization generating functional redundancy in the Ras/Rac pathway.
    Mol Biol Cell. 2004 Jan;15(1):91-8 PMID: 14565974
  42. Muscleblind-like 1 suppresses breast cancer metastatic colonization and stabilizes metastasis suppressor transcripts.
    Genes Dev. 2016 Feb 15;30(4):386-98 PMID: 26883358
  43. Ubiquitin-conjugating enzyme Ubc13 controls breast cancer metastasis through a TAK1-p38 MAP kinase cascade.
    Proc Natl Acad Sci U S A. 2014 Sep 23;111(38):13870-5 PMID: 25189770
  44. EZH2 inhibition decreases p38 signaling and suppresses breast cancer motility and metastasis.
    Breast Cancer Res Treat. 2013 Apr;138(3):741-52 PMID: 23539298
  45. Vinculin phosphorylation at residues Y100 and Y1065 is required for cellular force transmission.
    J Cell Sci. 2015 Sep 15;128(18):3435-43 PMID: 26240176
  46. The methyltransferase EZH2 is not required for mammary cancer development, although high EZH2 and low H3K27me3 correlate with poor prognosis of ER-positive breast cancers.
    Mol Carcinog. 2015 Oct;54(10):1172-80 PMID: 25043748
  47. The Stress Kinase p38α as a Target for Cancer Therapy.
    Cancer Res. 2015 Oct 1;75(19):3997-4002 PMID: 26377941
  48. CD24+ Ovarian Cancer Cells Are Enriched for Cancer-Initiating Cells and Dependent on JAK2 Signaling for Growth and Metastasis.
    Mol Cancer Ther. 2015 Jul;14(7):1717-27 PMID: 25969154
  49. Signal integration by JNK and p38 MAPK pathways in cancer development.
    Nat Rev Cancer. 2009 Aug;9(8):537-49 PMID: 19629069
  50. The phosphorylation of vinculin on tyrosine residues 100 and 1065, mediated by SRC kinases, affects cell spreading.
    Mol Biol Cell. 2004 Sep;15(9):4234-47 PMID: 15229287
  51. Loss of vinculin and membrane-bound β-catenin promotes metastasis and predicts poor prognosis in colorectal cancer.
    Mol Cancer. 2014 Dec 11;13:263 PMID: 25496021
  52. Phosphorylation primes vinculin for activation.
    Biophys J. 2012 May 2;102(9):2022-30 PMID: 22824265
Article Info
Journal
Nature communications
Abbr.
Nat Commun
ISSN
2041-1723
DOI
10.1038/s41467-018-05078-8
Email
kleer@umich.edu
Published
2018-00-18
Epub
2018-00-18
Pages
2801
Language
English
Region
England
NLM ID
101528555
PMCID
PMC6051995
Subset
IM
Grants
NCI NIH HHS · R01 CA125577 · United States
NCI NIH HHS · P30 CA046592 · United States
U.S. Department of Health &amp
Human Services · NIH · National Cancer Institute (NCI)
NCI NIH HHS · U24 CA210967 · United States
NIGMS NIH HHS · T32 GM007863 · United States
U.S. Department of Health &amp
Human Services · NIH · National Institute of General Medical Sciences (NIGMS)
NIGMS NIH HHS · R25 GM086262 · United States
NIGMS NIH HHS · R01 GM094231 · United States
NCI NIH HHS · R01 CA107469 · United States
U.S. Department of Health &amp
Human Services · NIH · National Cancer Institute (NCI)
U.S. Department of Health &amp
Human Services · NIH · National Cancer Institute (NCI)
NCI NIH HHS · T32 CA140044 · United States
U.S. Department of Health &amp
Human Services · NIH · National Institute of General Medical Sciences (NIGMS)
External Links
PubMed source DOI: 10.1038/s41467-018-05078-8 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2