Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels
Abstract Embryonic stem cell (ESC) differentiation and somatic cell reprogramming are biological processes governed by antagonistic expression or repression of a largely common set of genes. Accurate regulation of gene expression is thus essential for both processes, and alterations in RNA processin...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2021-06-01
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| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-021-03906-2 |
| _version_ | 1818928253315842048 |
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| author | Agnes Fütterer Amaia Talavera-Gutiérrez Tirso Pons Jesús de Celis Julio Gutiérrez Verónica Domínguez Plaza Carlos Martínez-A |
| author_facet | Agnes Fütterer Amaia Talavera-Gutiérrez Tirso Pons Jesús de Celis Julio Gutiérrez Verónica Domínguez Plaza Carlos Martínez-A |
| author_sort | Agnes Fütterer |
| collection | DOAJ |
| description | Abstract Embryonic stem cell (ESC) differentiation and somatic cell reprogramming are biological processes governed by antagonistic expression or repression of a largely common set of genes. Accurate regulation of gene expression is thus essential for both processes, and alterations in RNA processing are predicted to negatively affect both. We show that truncation of the DIDO gene alters RNA splicing and transcription termination in ESC and mouse embryo fibroblasts (MEF), which affects genes involved in both differentiation and reprogramming. We combined transcriptomic, protein interaction, and cellular studies to identify the underlying molecular mechanism. We found that DIDO3 interacts with the helicase DHX9, which is involved in R-loop processing and transcription termination, and that DIDO3-exon16 deletion increases nuclear R-loop content and causes DNA replication stress. Overall, these defects result in failure of ESC to differentiate and of MEF to be reprogrammed. MEF immortalization restored impaired reprogramming capacity. We conclude that DIDO3 has essential functions in ESC differentiation and somatic cell reprogramming by supporting accurate RNA metabolism, with its exon16-encoded domain playing the main role. |
| first_indexed | 2024-12-20T03:25:58Z |
| format | Article |
| id | doaj.art-e551a107dc534ddebf5373519f8ca162 |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-12-20T03:25:58Z |
| publishDate | 2021-06-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-e551a107dc534ddebf5373519f8ca1622022-12-21T19:55:05ZengNature Publishing GroupCell Death and Disease2041-48892021-06-0112711410.1038/s41419-021-03906-2Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levelsAgnes Fütterer0Amaia Talavera-Gutiérrez1Tirso Pons2Jesús de Celis3Julio Gutiérrez4Verónica Domínguez Plaza5Carlos Martínez-A6Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC)Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC)Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC)Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC)Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC)Transgenesis Unit, CNB & Centro de Biología Molecular Severo Ochoa, CSIC-Universidad Autónoma de MadridDepartment of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC)Abstract Embryonic stem cell (ESC) differentiation and somatic cell reprogramming are biological processes governed by antagonistic expression or repression of a largely common set of genes. Accurate regulation of gene expression is thus essential for both processes, and alterations in RNA processing are predicted to negatively affect both. We show that truncation of the DIDO gene alters RNA splicing and transcription termination in ESC and mouse embryo fibroblasts (MEF), which affects genes involved in both differentiation and reprogramming. We combined transcriptomic, protein interaction, and cellular studies to identify the underlying molecular mechanism. We found that DIDO3 interacts with the helicase DHX9, which is involved in R-loop processing and transcription termination, and that DIDO3-exon16 deletion increases nuclear R-loop content and causes DNA replication stress. Overall, these defects result in failure of ESC to differentiate and of MEF to be reprogrammed. MEF immortalization restored impaired reprogramming capacity. We conclude that DIDO3 has essential functions in ESC differentiation and somatic cell reprogramming by supporting accurate RNA metabolism, with its exon16-encoded domain playing the main role.https://doi.org/10.1038/s41419-021-03906-2 |
| spellingShingle | Agnes Fütterer Amaia Talavera-Gutiérrez Tirso Pons Jesús de Celis Julio Gutiérrez Verónica Domínguez Plaza Carlos Martínez-A Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels Cell Death and Disease |
| title | Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels |
| title_full | Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels |
| title_fullStr | Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels |
| title_full_unstemmed | Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels |
| title_short | Impaired stem cell differentiation and somatic cell reprogramming in DIDO3 mutants with altered RNA processing and increased R-loop levels |
| title_sort | impaired stem cell differentiation and somatic cell reprogramming in dido3 mutants with altered rna processing and increased r loop levels |
| url | https://doi.org/10.1038/s41419-021-03906-2 |
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