DOT1L methyltransferase regulates the calcium influx in erythroid progenitor cells in response to erythropoietin

Feng, Yi; Borosha, Shaon; Ratri, Anamika; Housami, Sami M.; Chakravarthi, V. Praveen; Wang, Huizhen; Vivian, Jay L.; Fields, Timothy A.; Kinsey, William H.; Rumi, MA Karim; Fields, Patrick E.

doi:10.1101/2020.10.04.325746

Cited by 2 publications

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References 21 publications

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“…We previously reported that hematopoietic transcription factor (TF) Gata2 was significantly reduced in Dot1L- KO HPCs, whereas Pu.1 , an erythropoiesis inhibiting TF was upregulated ( Feng et al, 2010 ). We also observed that KIT-positive HPCs from Dot1L -KO yolk sacs expressed low levels of Trpc6 ( Feng et al, 20202020 ). Our RNA-seq data recapitulated the previous observations regarding expression of Gata2, Pu.1 and Trpc6 (PRJNA666736).…”

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confidence: 52%

DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts

et al. 2022

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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.

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Section: Discussionsupporting

confidence: 52%

DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts

et al. 2022

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DOT1L primarily acts as a transcriptional repressor in hematopoietic progenitor cells

Borosha

Ratri

Housami

et al. 2020

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DOT1L is essential for early hematopoiesis but the precise mechanisms remain largely unclear. The only known function of DOT1L is histone H3 lysine 79 (H3K79) methylation. We generated two mouse models; a Dot1L-knockout (Dot1L-KO), and another possessing a point mutation in its methyltransferase domain (Dot1L-MM) to determine the role of its catalytic activity during early hematopoiesis. We observed that Dot1L-KO embryos suffered from severe anemia, while Dot1L-MM embryos showed minimal to no anemia. However, ex vivo culture of Dot1L-MM hematopoietic progenitors (HPCs) exhibited defective development of myeloid and mixed progenitors. DOT1L is a well-recognized, cell-type specific epigenetic regulator of gene expression. To elucidate the mechanisms underlying such diverse hematopoietic properties of Dot1L-KO and Dot1L-MM HPCs, we examined their whole transcriptomes. Extensively self-renewing erythroblast (ESRE) cultures were established using yolk sac (YS) cells collected on embryonic day 10.5 (E10.5). Dot1l-KO and Dot1l-MM cells expanded significantly less than the wildtype cells and showed slower progression through the cell cycle. Total RNA extracted from the wildtype and Dot1l-mutant ESRE cells were subjected to RNA-seq analyses. We observed that the majority (~82%) of the differentially expressed genes (DEGs) were upregulated in both of the Dot1L-mutants, which suggests that DOT1L predominantly acts as a transcriptional repressor in HPCs. We also observed that about ~40% of the DEGs were unique to either of the mutant group, suggesting that DOT1L possesses both methyltransferase domain-dependent and -independent functions. We further analyzed Gene Ontology and signaling pathways relevant to the DEGs common to both mutant groups and those that were unique to either group. Among the common DEGs, we observed upregulation of CDK inhibitors, which explains the cell cycle arrest in both of the Dot1L-mutant progenitors.

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DOT1L methyltransferase regulates the calcium influx in erythroid progenitor cells in response to erythropoietin

Cited by 2 publications

References 21 publications

DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts

DOT1L Mediated Gene Repression in Extensively Self-Renewing Erythroblasts

DOT1L primarily acts as a transcriptional repressor in hematopoietic progenitor cells

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