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Updated: Jul 9, 2025
Analysis of Cardiac Chamber Development During Mouse Embryogenesis Using Whole Mount Epifluorescence
Published on: April 17, 2019
The H2Bub1-deposition complex is required for human and mouse cardiogenesis
Syndi Barish1, Kathryn Berg1, Jeffrey Drozd2
1Department of Genetics, Yale University School of Medicine, 333 Cedar Street, New Haven, CT 06510, USA.
Insights
Histone H2B monoubiquitylation (H2Bub1) is crucial for heart development and cardiomyocyte differentiation. Proper H2Bub1 distribution ensures correct expression of tissue-specific genes, impacting cardiogenesis.
Area of Science:
- Epigenetics
- Developmental Biology
- Cardiovascular Research
Background:
- De novo variants in histone H2B monoubiquitylation (H2Bub1) are linked to congenital heart disease.
- H2Bub1 plays roles in stem cell differentiation, cilia function, cardiomyocyte maturation, and transcriptional elongation.
- The precise role of H2Bub1 in cardiogenesis remains unclear.
Purpose of the Study:
- To investigate the role of H2Bub1 and its deposition complex in mouse cardiogenesis and human cardiomyocyte differentiation.
- To elucidate how H2Bub1 distribution affects gene expression during cardiomyocyte development.
Main Methods:
- Analysis of H2Bub1 marks during human induced pluripotent stem cell (iPSC) differentiation into cardiomyocytes.
- Generation of mice with cardiac-specific Rnf20 deletion to assess its role in cardiogenesis.
- Quantification of full-length transcripts for cardiac-specific genes in iPSC-derived cardiomyocytes with altered H2Bub1 levels.
Main Results:
- The H2Bub1 deposition complex (RNF20-RNF40-UBE2B) is essential for mouse cardiogenesis and human iPSC-cardiomyocyte differentiation.
- Cardiac-specific Rnf20 deletion in mice leads to embryonic lethality and myocardial abnormalities.
- H2Bub1 is dynamically regulated during cardiomyocyte differentiation, preserved on key cardiac-specific genes, and its reduction impairs full-length transcript production.
Conclusions:
- Normal H2Bub1 distribution is indispensable for proper cardiogenesis and cardiomyocyte differentiation.
- H2Bub1 regulates tissue-specific gene expression, likely by promoting the production of full-length transcripts.
Abstract:
De novo variants affecting monoubiquitylation of histone H2B (H2Bub1) are enriched in human congenital heart disease. H2Bub1 is required in stem cell differentiation, cilia function, post-natal cardiomyocyte maturation and transcriptional elongation. However, how H2Bub1 affects cardiogenesis is unknown. We show that the H2Bub1-deposition complex (RNF20-RNF40-UBE2B) is required for mouse cardiogenesis and for differentiation of human iPSCs into cardiomyocytes. Mice with cardiac-specific Rnf20 deletion are embryonic lethal and have abnormal myocardium. We then analyzed H2Bub1 marks during differentiation of human iPSCs into cardiomyocytes. H2Bub1 is erased from most genes at the transition from cardiac mesoderm to cardiac progenitor cells but is preserved on a subset of long cardiac-specific genes. When H2Bub1 is reduced in iPSC-derived cardiomyocytes, long cardiac-specific genes have fewer full-length transcripts. This correlates with H2Bub1 accumulation near the center of these genes. H2Bub1 accumulation near the center of tissue-specific genes was also observed in embryonic fibroblasts and fetal osteoblasts. In summary, we show that normal H2Bub1 distribution is required for cardiogenesis and cardiomyocyte differentiation, and suggest that H2Bub1 regulates tissue-specific gene expression by increasing the amount of full-length transcripts.
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