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DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts
DOT1L is essential for embryonic hematopoiesis but the precise mechanisms of its action remain unclear. The only recognized function of DOT1L is histone H3 lysine 79 (H3K79) methylation, which has been implicated in both transcriptional activation and repression. We observed that deletion of the mou...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8984088/ https://www.ncbi.nlm.nih.gov/pubmed/35401699 http://dx.doi.org/10.3389/fgene.2022.828086 |
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author | Borosha, Shaon Ratri, Anamika Ghosh, Subhra Malcom, Carrie A. Chakravarthi, V. Praveen Vivian, Jay L. Fields, Timothy A. Rumi, M. A. Karim Fields, Patrick E. |
author_facet | Borosha, Shaon Ratri, Anamika Ghosh, Subhra Malcom, Carrie A. Chakravarthi, V. Praveen Vivian, Jay L. Fields, Timothy A. Rumi, M. A. Karim Fields, Patrick E. |
author_sort | Borosha, Shaon |
collection | PubMed |
description | DOT1L is essential for embryonic hematopoiesis but the precise mechanisms of its action remain unclear. The only recognized function of DOT1L is histone H3 lysine 79 (H3K79) methylation, which has been implicated in both transcriptional activation and repression. We observed that deletion of the mouse Dot1L gene (Dot1L-KO) or selective mutation of its methyltransferase domain (Dot1L-MM) can differentially affect early embryonic erythropoiesis. However, both mutations result in embryonic lethality by mid-gestation and growth of hematopoietic progenitor cells (HPCs) is similarly affected in extensively self-renewing erythroblast (ESRE) cultures established from yolk sac cells. To understand DOT1L-mediated gene regulation and to clarify the role of H3K79 methylation, we analyzed whole transcriptomes of wildtype and Dot1L-mutant ESRE cells. We observed that more than 80% of the differentially expressed genes (DEGs) were upregulated in the mutant ESRE cells either lacking the DOT1L protein or the DOT1L methyltransferase activity. However, approximately 45% of the DEGs were unique to either mutant group, indicating that DOT1L possesses both methyltransferase-dependent and -independent gene regulatory functions. Analyses of Gene Ontology and signaling pathways for the DEGs were consistent, with DEGs that were found to be common or unique to either mutant group. Genes related to proliferation of HPCs were primarily impacted in Dot1L-KO cells, while genes related to HPC development were affected in the Dot1L-MM cells. A subset of genes related to differentiation of HPCs were affected in both mutant groups of ESREs. Our findings suggest that DOT1L primarily acts to repress gene expression in HPCs, and this function can be independent of its methyltransferase activity. |
format | Online Article Text |
id | pubmed-8984088 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-89840882022-04-07 DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts Borosha, Shaon Ratri, Anamika Ghosh, Subhra Malcom, Carrie A. Chakravarthi, V. Praveen Vivian, Jay L. Fields, Timothy A. Rumi, M. A. Karim Fields, Patrick E. Front Genet Genetics DOT1L is essential for embryonic hematopoiesis but the precise mechanisms of its action remain unclear. The only recognized function of DOT1L is histone H3 lysine 79 (H3K79) methylation, which has been implicated in both transcriptional activation and repression. We observed that deletion of the mouse Dot1L gene (Dot1L-KO) or selective mutation of its methyltransferase domain (Dot1L-MM) can differentially affect early embryonic erythropoiesis. However, both mutations result in embryonic lethality by mid-gestation and growth of hematopoietic progenitor cells (HPCs) is similarly affected in extensively self-renewing erythroblast (ESRE) cultures established from yolk sac cells. To understand DOT1L-mediated gene regulation and to clarify the role of H3K79 methylation, we analyzed whole transcriptomes of wildtype and Dot1L-mutant ESRE cells. We observed that more than 80% of the differentially expressed genes (DEGs) were upregulated in the mutant ESRE cells either lacking the DOT1L protein or the DOT1L methyltransferase activity. However, approximately 45% of the DEGs were unique to either mutant group, indicating that DOT1L possesses both methyltransferase-dependent and -independent gene regulatory functions. Analyses of Gene Ontology and signaling pathways for the DEGs were consistent, with DEGs that were found to be common or unique to either mutant group. Genes related to proliferation of HPCs were primarily impacted in Dot1L-KO cells, while genes related to HPC development were affected in the Dot1L-MM cells. A subset of genes related to differentiation of HPCs were affected in both mutant groups of ESREs. Our findings suggest that DOT1L primarily acts to repress gene expression in HPCs, and this function can be independent of its methyltransferase activity. Frontiers Media S.A. 2022-03-23 /pmc/articles/PMC8984088/ /pubmed/35401699 http://dx.doi.org/10.3389/fgene.2022.828086 Text en Copyright © 2022 Borosha, Ratri, Ghosh, Malcom, Chakravarthi, Vivian, Fields, Rumi and Fields. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Genetics Borosha, Shaon Ratri, Anamika Ghosh, Subhra Malcom, Carrie A. Chakravarthi, V. Praveen Vivian, Jay L. Fields, Timothy A. Rumi, M. A. Karim Fields, Patrick E. DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts |
title | DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts |
title_full | DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts |
title_fullStr | DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts |
title_full_unstemmed | DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts |
title_short | DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts |
title_sort | dot1l mediated gene repression in extensively self-renewing erythroblasts |
topic | Genetics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8984088/ https://www.ncbi.nlm.nih.gov/pubmed/35401699 http://dx.doi.org/10.3389/fgene.2022.828086 |
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