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Keywords {🔍}

cells, nsun, mitochondrial, pubmed, escs, cell, article, differentiation, google, scholar, wildtype, fig, cas, stem, rna, levels, catcat, nsunmutant, ros, central, translation, expression, methylation, human, wnt, function, mttrnamet, embryonic, esc, protein, shown, proliferation, reduced, mouse, alkbh, performed, cytosine, mutant, pluripotency, medium, test, potential, trna, modification, increased, methyltransferase, modifications, found, mitochondria, glycolysis,

Topics {✒️}

replication marker proliferating-cell-nuclear-antigen article download pdf wnt/beta-catenin signaling pathway c-terminal egfp tag 35s-pulse-labelled mitochondrial proteins mitotracker red cm-h2xros real-time pcr-based determination strand-specific ssrna probes single-cell transcriptomics show ice-cold ripa buffer nsun3 cat/cat compared nsun3 cat/cat cells molecular biology nsun3 cat/cat ebs mterf4 regulates translation nsun3 cat/cat escs wild-type stem cells nsun3 cat/cat relative 50 mm na-phosphate buffer mediates myc-induced proliferation wild-type cells indicating functional peptidyl-trna hydrolase nsun3-mutant cells correlated full size image employed crispr/cas9 technology serum-free dmem medium pcr-mediated bisulfite sequencing embryonic stem cells nsun3-mutant es cells wild-type es cells nsun3-mutant cells compared ribosome-based quality control reach wild-type levels catalytically inactive nsun3 cell line expressing catalytically inactive mutant nsun3-mutant ebs compared strand-specific probes pluripotent stem cells px-nsun3-gfp plasmid nsun3-mutant escs compared promoting mitochondrial activity full access gelatine-coated culture dishes rna-binding protein required nsun3 cat/cat mt-trnamet wobble cytosine cell cycle profile stem cell pluripotency austrian science fund

Questions {❓}

• Sherwood V (2015) WNT signaling: an emerging mediator of cancer cell metabolism?

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         headline:RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
         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 tRNAMet. 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-tRNAMet 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.
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      headline:RNA cytosine methyltransferase Nsun3 regulates embryonic stem cell differentiation by promoting mitochondrial activity
      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 tRNAMet. 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-tRNAMet 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.
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            type:Organization
      name:Jakob Troppmair
      affiliation:
            name:Medical University of Innsbruck
            address:
               name:Daniel Swarovski Research Laboratory, Department of Visceral, Transplant, and Thoracic Surgery, Medical University of Innsbruck, Innsbruck, Austria
               type:PostalAddress
            type:Organization
      name:Alexandra Lusser
      url:http://orcid.org/0000-0002-2226-9081
      affiliation:
            name:Medical University of Innsbruck
            address:
               name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
               type:PostalAddress
            type:Organization
      email:[email protected]
PostalAddress:
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Daniel Swarovski Research Laboratory, Department of Visceral, Transplant, and Thoracic Surgery, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria
      name:Zymo Research Corp., Irvine, USA
      name:Institute for Neuroscience, Medical University of Innsbruck, Innsbruck, Austria
      name:Daniel Swarovski Research Laboratory, Department of Visceral, Transplant, and Thoracic Surgery, Medical University of Innsbruck, Innsbruck, Austria
      name:Division of Molecular Biology, Biocenter, Medical University of Innsbruck, Innsbruck, Austria

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