PreprintArticleVersion 1Preserved in Portico This version is not peer-reviewed
Epigenetic Reprogramming and Inheritance of the Cellular Differentiation Status Following Transient Expression of a Nonfunctional DN Rb Mutant in Murine MSCs
Version 1
: Received: 4 September 2024 / Approved: 4 September 2024 / Online: 5 September 2024 (07:12:27 CEST)
How to cite:
Mikhail, B.; Maksimova, I.; Sasoveca, I. Epigenetic Reprogramming and Inheritance of the Cellular Differentiation Status Following Transient Expression of a Nonfunctional DN Rb Mutant in Murine MSCs. Preprints2024, 2024090375. https://doi.org/10.20944/preprints202409.0375.v1
Mikhail, B.; Maksimova, I.; Sasoveca, I. Epigenetic Reprogramming and Inheritance of the Cellular Differentiation Status Following Transient Expression of a Nonfunctional DN Rb Mutant in Murine MSCs. Preprints 2024, 2024090375. https://doi.org/10.20944/preprints202409.0375.v1
Mikhail, B.; Maksimova, I.; Sasoveca, I. Epigenetic Reprogramming and Inheritance of the Cellular Differentiation Status Following Transient Expression of a Nonfunctional DN Rb Mutant in Murine MSCs. Preprints2024, 2024090375. https://doi.org/10.20944/preprints202409.0375.v1
APA Style
Mikhail, B., Maksimova, I., & Sasoveca, I. (2024). Epigenetic Reprogramming and Inheritance of the Cellular Differentiation Status Following Transient Expression of a Nonfunctional DN Rb Mutant in Murine MSCs. Preprints. https://doi.org/10.20944/preprints202409.0375.v1
Chicago/Turabian Style
Mikhail, B., Irina Maksimova and Ilona Sasoveca. 2024 "Epigenetic Reprogramming and Inheritance of the Cellular Differentiation Status Following Transient Expression of a Nonfunctional DN Rb Mutant in Murine MSCs" Preprints. https://doi.org/10.20944/preprints202409.0375.v1
Abstract
The retinoblastoma gene product (Rb1), a master regulator of the cell cycle, also plays a prominent role in cell differentiation. Previously, by analyzing the differentiation of cells transiently overexpressing the ΔS/N DN Rb1 mutant, we demonstrated that these cells fail to differentiate into mature adipocytes and that they constitutively silence Pparγ2 through CpG methylation. Here, we demonstrate that the consequences of transient expression of ΔS/N DN Rb1 are accompanied by retention of Cebpa promoter methylation near the TSS under adipogenic differentiation, thereby preventing its expression. The CGIs of the promoters of the Rb1, Ezh2, Mll4, Utx and Tet2 genes, which are essential for adipogenic differentiation, have an unmethylated status regardless of the cell differentiation state. Moreover, Dnmt3a, a de novo DNA methyltransferase, is overexpressed in undifferentiated ΔS/N cells compared with wild-type cells, and in addition to Dnmt1, Dnmt3a is significantly upregulated by adipogenic stimuli in both wild-type and ΔS/N cells. Notably, the chromatin modifier Ezh2, which is also involved in epigenetic reprogramming, is highly induced in ΔS/N cells. Overall, we demonstrate that two major genes, Pparγ2 and Cebpa, which are responsible for terminal adipocyte differentiation, are selectively epigenetically reprogrammed to constitutively silent states. We hypothesize that the activation of Dnmt3a, Rb1 and Ezh2 observed in ΔS/N cells may be a consequence of a stress response caused by the accumulation and malfunctioning of Rb1-interacting complexes for epigenetic reprogramming of Pparγ2/Cebpa and prevention of adipogenesis in an inappropriate cellular context. The failure of ΔS/N cells to differentiate and express Pparγ2 and Cebpa in culture following the expression of the DN Rb1 mutant may indicate the creation of epigenetic memory for new reprogrammed epigenetic states of genes.
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
