Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes
Lsh is a chromatin remodeling factor that regulates DNA methylation and chromatin function in mammals. The dynamics of these chromatin changes and whether they are directly controlled by Lsh remain unclear. To understand the molecular mechanisms of Lsh chromatin controlled regulation of gene express...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2018-02-01
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| Series: | Epigenetics |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/15592294.2017.1338234 |
| _version_ | 1797678992254304256 |
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| author | Jianke Ren Nathaniel A. Hathaway Gerald R. Crabtree Kathrin Muegge |
| author_facet | Jianke Ren Nathaniel A. Hathaway Gerald R. Crabtree Kathrin Muegge |
| author_sort | Jianke Ren |
| collection | DOAJ |
| description | Lsh is a chromatin remodeling factor that regulates DNA methylation and chromatin function in mammals. The dynamics of these chromatin changes and whether they are directly controlled by Lsh remain unclear. To understand the molecular mechanisms of Lsh chromatin controlled regulation of gene expression, we established a tethering system that recruits a Gal4-Lsh fusion protein to an engineered Oct4 locus through Gal4 binding sites in murine embryonic stem (ES) cells. We examined the molecular epigenetic events induced by Lsh binding including: histone modification, DNA methylation and chromatin accessibility to determine nucleosome occupancy before and after embryonic stem cell differentiation. Our results indicate that Lsh assists gene repression upon binding to the Oct4 promoter region. Furthermore, we detected less chromatin accessibility and reduced active histone modifications at the tethered site in undifferentiated ES, while GFP reporter gene expression and DNA methylation patterns remained unchanged at this stage. Upon differentiation, association of Lsh promotes transcriptional repression of the reporter gene accompanied by the increase of repressive histone marks and a gain of DNA methylation at distal and proximal Oct4 enhancer sites. Taken together, this approach allowed us to examine Lsh mediated epigenetic regulation as a dynamic process and revealed chromatin accessibility changes as the primary consequence of Lsh function. |
| first_indexed | 2024-03-11T23:07:56Z |
| format | Article |
| id | doaj.art-f0eda58b559340a0b4fdc6a575af760b |
| institution | Directory Open Access Journal |
| issn | 1559-2294 1559-2308 |
| language | English |
| last_indexed | 2024-03-11T23:07:56Z |
| publishDate | 2018-02-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Epigenetics |
| spelling | doaj.art-f0eda58b559340a0b4fdc6a575af760b2023-09-21T12:43:13ZengTaylor & Francis GroupEpigenetics1559-22941559-23082018-02-0113217318110.1080/15592294.2017.13382341338234Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changesJianke Ren0Nathaniel A. Hathaway1Gerald R. Crabtree2Kathrin Muegge3Center for Cancer Research, National Cancer InstituteUNC Eshelman School of PharmacyStanford University School of Medicine, Howard Hughes Medical InstituteCenter for Cancer Research, National Cancer InstituteLsh is a chromatin remodeling factor that regulates DNA methylation and chromatin function in mammals. The dynamics of these chromatin changes and whether they are directly controlled by Lsh remain unclear. To understand the molecular mechanisms of Lsh chromatin controlled regulation of gene expression, we established a tethering system that recruits a Gal4-Lsh fusion protein to an engineered Oct4 locus through Gal4 binding sites in murine embryonic stem (ES) cells. We examined the molecular epigenetic events induced by Lsh binding including: histone modification, DNA methylation and chromatin accessibility to determine nucleosome occupancy before and after embryonic stem cell differentiation. Our results indicate that Lsh assists gene repression upon binding to the Oct4 promoter region. Furthermore, we detected less chromatin accessibility and reduced active histone modifications at the tethered site in undifferentiated ES, while GFP reporter gene expression and DNA methylation patterns remained unchanged at this stage. Upon differentiation, association of Lsh promotes transcriptional repression of the reporter gene accompanied by the increase of repressive histone marks and a gain of DNA methylation at distal and proximal Oct4 enhancer sites. Taken together, this approach allowed us to examine Lsh mediated epigenetic regulation as a dynamic process and revealed chromatin accessibility changes as the primary consequence of Lsh function.http://dx.doi.org/10.1080/15592294.2017.1338234dna methylationepigeneticshistone modificationsnucleosome occupancytethering |
| spellingShingle | Jianke Ren Nathaniel A. Hathaway Gerald R. Crabtree Kathrin Muegge Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes Epigenetics dna methylation epigenetics histone modifications nucleosome occupancy tethering |
| title | Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes |
| title_full | Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes |
| title_fullStr | Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes |
| title_full_unstemmed | Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes |
| title_short | Tethering of Lsh at the Oct4 locus promotes gene repression associated with epigenetic changes |
| title_sort | tethering of lsh at the oct4 locus promotes gene repression associated with epigenetic changes |
| topic | dna methylation epigenetics histone modifications nucleosome occupancy tethering |
| url | http://dx.doi.org/10.1080/15592294.2017.1338234 |
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