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RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity

Chemical modifications of RNA have been attracting increasing interest because of their impact on RNA fate and function. Therefore, the characterization of enzymes catalyzing such modifications is of great importance. The RNA cytosine methyltransferase NSUN3 was recently shown to generate 5-methylcy...

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Autores principales: Trixl, Lukas, Amort, Thomas, Wille, Alexandra, Zinni, Manuela, Ebner, Susanne, Hechenberger, Clara, Eichin, Felix, Gabriel, Hanna, Schoberleitner, Ines, Huang, Anming, Piatti, Paolo, Nat, Roxana, Troppmair, Jakob, Lusser, Alexandra
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer International Publishing 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5852174/
https://www.ncbi.nlm.nih.gov/pubmed/29103146
http://dx.doi.org/10.1007/s00018-017-2700-0
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author Trixl, Lukas
Amort, Thomas
Wille, Alexandra
Zinni, Manuela
Ebner, Susanne
Hechenberger, Clara
Eichin, Felix
Gabriel, Hanna
Schoberleitner, Ines
Huang, Anming
Piatti, Paolo
Nat, Roxana
Troppmair, Jakob
Lusser, Alexandra
author_facet Trixl, Lukas
Amort, Thomas
Wille, Alexandra
Zinni, Manuela
Ebner, Susanne
Hechenberger, Clara
Eichin, Felix
Gabriel, Hanna
Schoberleitner, Ines
Huang, Anming
Piatti, Paolo
Nat, Roxana
Troppmair, Jakob
Lusser, Alexandra
author_sort Trixl, Lukas
collection PubMed
description Chemical modifications of RNA have been attracting increasing interest because of their impact on RNA fate and function. Therefore, the characterization of enzymes catalyzing such modifications is of great importance. The RNA cytosine methyltransferase NSUN3 was recently shown to generate 5-methylcytosine in the anticodon loop of mitochondrial tRNA(Met). Further oxidation of this position is required for normal mitochondrial translation and function in human somatic cells. Because embryonic stem cells (ESCs) are less dependent on oxidative phosphorylation than somatic cells, we examined the effects of catalytic inactivation of Nsun3 on self-renewal and differentiation potential of murine ESCs. We demonstrate that Nsun3-mutant cells show strongly reduced mt-tRNA(Met) methylation and formylation as well as reduced mitochondrial translation and respiration. Despite the lower dependence of ESCs on mitochondrial activity, proliferation of mutant cells was reduced, while pluripotency marker gene expression was not affected. By contrast, ESC differentiation was skewed towards the meso- and endoderm lineages at the expense of neuroectoderm. Wnt3 was overexpressed in early differentiating mutant embryoid bodies and in ESCs, suggesting that impaired mitochondrial function disturbs normal differentiation programs by interfering with cellular signalling pathways. Interestingly, basal levels of reactive oxygen species (ROS) were not altered in ESCs, but Nsun3 inactivation attenuated induction of mitochondrial ROS upon stress, which may affect gene expression programs upon differentiation. Our findings not only characterize Nsun3 as an important regulator of stem cell fate but also provide a model system to study the still incompletely understood interplay of mitochondrial function with stem cell pluripotency and differentiation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00018-017-2700-0) contains supplementary material, which is available to authorized users.
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spelling pubmed-58521742018-03-21 RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity Trixl, Lukas Amort, Thomas Wille, Alexandra Zinni, Manuela Ebner, Susanne Hechenberger, Clara Eichin, Felix Gabriel, Hanna Schoberleitner, Ines Huang, Anming Piatti, Paolo Nat, Roxana Troppmair, Jakob Lusser, Alexandra Cell Mol Life Sci Original Article Chemical modifications of RNA have been attracting increasing interest because of their impact on RNA fate and function. Therefore, the characterization of enzymes catalyzing such modifications is of great importance. The RNA cytosine methyltransferase NSUN3 was recently shown to generate 5-methylcytosine in the anticodon loop of mitochondrial tRNA(Met). Further oxidation of this position is required for normal mitochondrial translation and function in human somatic cells. Because embryonic stem cells (ESCs) are less dependent on oxidative phosphorylation than somatic cells, we examined the effects of catalytic inactivation of Nsun3 on self-renewal and differentiation potential of murine ESCs. We demonstrate that Nsun3-mutant cells show strongly reduced mt-tRNA(Met) methylation and formylation as well as reduced mitochondrial translation and respiration. Despite the lower dependence of ESCs on mitochondrial activity, proliferation of mutant cells was reduced, while pluripotency marker gene expression was not affected. By contrast, ESC differentiation was skewed towards the meso- and endoderm lineages at the expense of neuroectoderm. Wnt3 was overexpressed in early differentiating mutant embryoid bodies and in ESCs, suggesting that impaired mitochondrial function disturbs normal differentiation programs by interfering with cellular signalling pathways. Interestingly, basal levels of reactive oxygen species (ROS) were not altered in ESCs, but Nsun3 inactivation attenuated induction of mitochondrial ROS upon stress, which may affect gene expression programs upon differentiation. Our findings not only characterize Nsun3 as an important regulator of stem cell fate but also provide a model system to study the still incompletely understood interplay of mitochondrial function with stem cell pluripotency and differentiation. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s00018-017-2700-0) contains supplementary material, which is available to authorized users. Springer International Publishing 2017-11-04 2018 /pmc/articles/PMC5852174/ /pubmed/29103146 http://dx.doi.org/10.1007/s00018-017-2700-0 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Trixl, Lukas
Amort, Thomas
Wille, Alexandra
Zinni, Manuela
Ebner, Susanne
Hechenberger, Clara
Eichin, Felix
Gabriel, Hanna
Schoberleitner, Ines
Huang, Anming
Piatti, Paolo
Nat, Roxana
Troppmair, Jakob
Lusser, Alexandra
RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
title RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
title_full RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
title_fullStr RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
title_full_unstemmed RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
title_short RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
title_sort rna cytosine methyltransferase nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5852174/
https://www.ncbi.nlm.nih.gov/pubmed/29103146
http://dx.doi.org/10.1007/s00018-017-2700-0
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