Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin
DNA methyltransferase I plays the central role in maintenance of CpG DNA methylation patterns across the genome and alteration of CpG methylation patterns is a frequent and significant occurrence across many cancers. Cancer cells carrying hypomorphic alleles of Dnmt1 have become important tools for...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2020-02-01
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| Series: | Epigenetics |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/15592294.2019.1656154 |
| _version_ | 1797678866247974912 |
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| author | Emily H Bowler Alex Smith-Vidal Alex Lester Joseph Bell Zhenghe Wang Christopher G. Bell Yihua Wang Nullin Divecha Paul J. Skipp Rob M. Ewing |
| author_facet | Emily H Bowler Alex Smith-Vidal Alex Lester Joseph Bell Zhenghe Wang Christopher G. Bell Yihua Wang Nullin Divecha Paul J. Skipp Rob M. Ewing |
| author_sort | Emily H Bowler |
| collection | DOAJ |
| description | DNA methyltransferase I plays the central role in maintenance of CpG DNA methylation patterns across the genome and alteration of CpG methylation patterns is a frequent and significant occurrence across many cancers. Cancer cells carrying hypomorphic alleles of Dnmt1 have become important tools for understanding Dnmt1 function and CpG methylation. In this study, we analyse colorectal cancer cells with a homozygous deletion of exons 3 to 5 of Dnmt1, resulting in reduced Dnmt1 activity. Although this cell model has been widely used to study the epigenome, the effects of the Dnmt1 hypomorph on cell signalling pathways and the wider proteome are largely unknown. In this study, we perform the first quantitative proteomic analysis of this important cell model and identify multiple signalling pathways and processes that are significantly dysregulated in the hypomorph cells. In Dnmt1 hypomorph cells, we observed a clear and unexpected signature of increased Epithelial-to-Mesenchymal transition (EMT) markers as well as reduced expression and sub-cellular re-localization of Beta-Catenin. Expression of wild-type Dnmt1 in hypomorph cells or knock-down of wild-type Dnmt1 did not recapitulate or rescue the observed protein profiles in Dnmt1 hypomorph cells suggesting that hypomorphic Dnmt1 causes changes not solely attributable to Dnmt1 protein levels. In summary, we present the first comprehensive proteomic analysis of the widely studied Dnmt1 hypomorph colorectal cancer cells and identify redistribution of Dnmt1 and its interaction partner Beta-Catenin. |
| first_indexed | 2024-03-11T23:06:09Z |
| format | Article |
| id | doaj.art-13527b518e204fa395e078462a3e3f77 |
| institution | Directory Open Access Journal |
| issn | 1559-2294 1559-2308 |
| language | English |
| last_indexed | 2024-03-11T23:06:09Z |
| publishDate | 2020-02-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Epigenetics |
| spelling | doaj.art-13527b518e204fa395e078462a3e3f772023-09-21T13:09:22ZengTaylor & Francis GroupEpigenetics1559-22941559-23082020-02-01151-210712110.1080/15592294.2019.16561541656154Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-CateninEmily H Bowler0Alex Smith-Vidal1Alex Lester2Joseph Bell3Zhenghe Wang4Christopher G. Bell5Yihua Wang6Nullin Divecha7Paul J. Skipp8Rob M. Ewing9University of SouthamptonUniversity of SouthamptonUniversity of SouthamptonUniversity of SouthamptonCase Western Reserve UniversityUniversity of SouthamptonUniversity of SouthamptonUniversity of SouthamptonUniversity of SouthamptonUniversity of SouthamptonDNA methyltransferase I plays the central role in maintenance of CpG DNA methylation patterns across the genome and alteration of CpG methylation patterns is a frequent and significant occurrence across many cancers. Cancer cells carrying hypomorphic alleles of Dnmt1 have become important tools for understanding Dnmt1 function and CpG methylation. In this study, we analyse colorectal cancer cells with a homozygous deletion of exons 3 to 5 of Dnmt1, resulting in reduced Dnmt1 activity. Although this cell model has been widely used to study the epigenome, the effects of the Dnmt1 hypomorph on cell signalling pathways and the wider proteome are largely unknown. In this study, we perform the first quantitative proteomic analysis of this important cell model and identify multiple signalling pathways and processes that are significantly dysregulated in the hypomorph cells. In Dnmt1 hypomorph cells, we observed a clear and unexpected signature of increased Epithelial-to-Mesenchymal transition (EMT) markers as well as reduced expression and sub-cellular re-localization of Beta-Catenin. Expression of wild-type Dnmt1 in hypomorph cells or knock-down of wild-type Dnmt1 did not recapitulate or rescue the observed protein profiles in Dnmt1 hypomorph cells suggesting that hypomorphic Dnmt1 causes changes not solely attributable to Dnmt1 protein levels. In summary, we present the first comprehensive proteomic analysis of the widely studied Dnmt1 hypomorph colorectal cancer cells and identify redistribution of Dnmt1 and its interaction partner Beta-Catenin.http://dx.doi.org/10.1080/15592294.2019.1656154dna methyltransferasebeta-cateninepithelial-mesenchymal transitionproteomics |
| spellingShingle | Emily H Bowler Alex Smith-Vidal Alex Lester Joseph Bell Zhenghe Wang Christopher G. Bell Yihua Wang Nullin Divecha Paul J. Skipp Rob M. Ewing Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin Epigenetics dna methyltransferase beta-catenin epithelial-mesenchymal transition proteomics |
| title | Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin |
| title_full | Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin |
| title_fullStr | Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin |
| title_full_unstemmed | Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin |
| title_short | Deep proteomic analysis of Dnmt1 mutant/hypomorphic colorectal cancer cells reveals dysregulation of epithelial–mesenchymal transition and subcellular re-localization of Beta-Catenin |
| title_sort | deep proteomic analysis of dnmt1 mutant hypomorphic colorectal cancer cells reveals dysregulation of epithelial mesenchymal transition and subcellular re localization of beta catenin |
| topic | dna methyltransferase beta-catenin epithelial-mesenchymal transition proteomics |
| url | http://dx.doi.org/10.1080/15592294.2019.1656154 |
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