Citrullination of HP1γ chromodomain affects association with chromatin
Abstract Background Stem cell differentiation involves major chromatin reorganisation, heterochromatin formation and genomic relocalisation of structural proteins, including heterochromatin protein 1 gamma (HP1γ). As the principal reader of the repressive histone marks H3K9me2/3, HP1 plays a key rol...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2019-04-01
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| Series: | Epigenetics & Chromatin |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s13072-019-0265-x |
| _version_ | 1818851797603713024 |
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| author | Meike Wiese Andrew J. Bannister Srinjan Basu Wayne Boucher Kai Wohlfahrt Maria A. Christophorou Michael L. Nielsen David Klenerman Ernest D. Laue Tony Kouzarides |
| author_facet | Meike Wiese Andrew J. Bannister Srinjan Basu Wayne Boucher Kai Wohlfahrt Maria A. Christophorou Michael L. Nielsen David Klenerman Ernest D. Laue Tony Kouzarides |
| author_sort | Meike Wiese |
| collection | DOAJ |
| description | Abstract Background Stem cell differentiation involves major chromatin reorganisation, heterochromatin formation and genomic relocalisation of structural proteins, including heterochromatin protein 1 gamma (HP1γ). As the principal reader of the repressive histone marks H3K9me2/3, HP1 plays a key role in numerous processes including heterochromatin formation and maintenance. Results We find that HP1γ is citrullinated in mouse embryonic stem cells (mESCs) and this diminishes when cells differentiate, indicating that it is a dynamically regulated post-translational modification during stem cell differentiation. Peptidylarginine deiminase 4, a known regulator of pluripotency, citrullinates HP1γ in vitro. This requires R38 and R39 within the HP1γ chromodomain, and the catalytic activity is enhanced by trimethylated H3K9 (H3K9me3) peptides. Mutation of R38 and R39, designed to mimic citrullination, affects HP1γ binding to H3K9me3-containing peptides. Using live-cell single-particle tracking, we demonstrate that R38 and R39 are important for HP1γ binding to chromatin in vivo. Furthermore, their mutation reduces the residence time of HP1γ on chromatin in differentiating mESCs. Conclusion Citrullination is a novel post-translational modification of the structural heterochromatin protein HP1γ in mESCs that is dynamically regulated during mESC differentiation. The citrullinated residues lie within the HP1γ chromodomain and are important for H3K9me3 binding in vitro and chromatin association in vivo. |
| first_indexed | 2024-12-19T07:10:44Z |
| format | Article |
| id | doaj.art-3abeefb38c8647619e3e0901093a5eca |
| institution | Directory Open Access Journal |
| issn | 1756-8935 |
| language | English |
| last_indexed | 2024-12-19T07:10:44Z |
| publishDate | 2019-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Epigenetics & Chromatin |
| spelling | doaj.art-3abeefb38c8647619e3e0901093a5eca2022-12-21T20:31:11ZengBMCEpigenetics & Chromatin1756-89352019-04-0112111710.1186/s13072-019-0265-xCitrullination of HP1γ chromodomain affects association with chromatinMeike Wiese0Andrew J. Bannister1Srinjan Basu2Wayne Boucher3Kai Wohlfahrt4Maria A. Christophorou5Michael L. Nielsen6David Klenerman7Ernest D. Laue8Tony Kouzarides9The Gurdon Institute, University of CambridgeThe Gurdon Institute, University of CambridgeDepartment of Biochemistry, University of CambridgeDepartment of Biochemistry, University of CambridgeDepartment of Biochemistry, University of CambridgeInstitute of Genetics and Molecular Medicine, Western General Hospital, University of EdinburghDepartment of Proteomics, The Novo Nordisk Foundation Center for Protein Research, Faculty of Health Sciences, University of CopenhagenDepartment of Chemistry, University of CambridgeDepartment of Biochemistry, University of CambridgeThe Gurdon Institute, University of CambridgeAbstract Background Stem cell differentiation involves major chromatin reorganisation, heterochromatin formation and genomic relocalisation of structural proteins, including heterochromatin protein 1 gamma (HP1γ). As the principal reader of the repressive histone marks H3K9me2/3, HP1 plays a key role in numerous processes including heterochromatin formation and maintenance. Results We find that HP1γ is citrullinated in mouse embryonic stem cells (mESCs) and this diminishes when cells differentiate, indicating that it is a dynamically regulated post-translational modification during stem cell differentiation. Peptidylarginine deiminase 4, a known regulator of pluripotency, citrullinates HP1γ in vitro. This requires R38 and R39 within the HP1γ chromodomain, and the catalytic activity is enhanced by trimethylated H3K9 (H3K9me3) peptides. Mutation of R38 and R39, designed to mimic citrullination, affects HP1γ binding to H3K9me3-containing peptides. Using live-cell single-particle tracking, we demonstrate that R38 and R39 are important for HP1γ binding to chromatin in vivo. Furthermore, their mutation reduces the residence time of HP1γ on chromatin in differentiating mESCs. Conclusion Citrullination is a novel post-translational modification of the structural heterochromatin protein HP1γ in mESCs that is dynamically regulated during mESC differentiation. The citrullinated residues lie within the HP1γ chromodomain and are important for H3K9me3 binding in vitro and chromatin association in vivo.http://link.springer.com/article/10.1186/s13072-019-0265-xChromatinCitrullinationStem cell differentiation |
| spellingShingle | Meike Wiese Andrew J. Bannister Srinjan Basu Wayne Boucher Kai Wohlfahrt Maria A. Christophorou Michael L. Nielsen David Klenerman Ernest D. Laue Tony Kouzarides Citrullination of HP1γ chromodomain affects association with chromatin Epigenetics & Chromatin Chromatin Citrullination Stem cell differentiation |
| title | Citrullination of HP1γ chromodomain affects association with chromatin |
| title_full | Citrullination of HP1γ chromodomain affects association with chromatin |
| title_fullStr | Citrullination of HP1γ chromodomain affects association with chromatin |
| title_full_unstemmed | Citrullination of HP1γ chromodomain affects association with chromatin |
| title_short | Citrullination of HP1γ chromodomain affects association with chromatin |
| title_sort | citrullination of hp1γ chromodomain affects association with chromatin |
| topic | Chromatin Citrullination Stem cell differentiation |
| url | http://link.springer.com/article/10.1186/s13072-019-0265-x |
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