The mitochondrial methylation potential gates mitoribosome assembly

Abstract S-adenosylmethionine (SAM) is the principal methyl donor in cells and is essential for mitochondrial gene expression, influencing RNA modifications, translation, and ribosome biogenesis. Using direct long-read RNA sequencing in mouse tissues and embryonic fibroblasts, we show that processin...

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Main Authors: Ruth I. C. Glasgow, Vivek Singh, Lucía Peña-Pérez, Alissa Wilhalm, Marco F. Moedas, David Moore, Florian A. Rosenberger, Xinping Li, Ilian Atanassov, Mira Saba, Miriam Cipullo, Joanna Rorbach, Anna Wedell, Christoph Freyer, Alexey Amunts, Anna Wredenberg
Format: Article
Language:English
Published: Nature Portfolio 2025-06-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-60977-x
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author Ruth I. C. Glasgow
Vivek Singh
Lucía Peña-Pérez
Alissa Wilhalm
Marco F. Moedas
David Moore
Florian A. Rosenberger
Xinping Li
Ilian Atanassov
Mira Saba
Miriam Cipullo
Joanna Rorbach
Anna Wedell
Christoph Freyer
Alexey Amunts
Anna Wredenberg
author_facet Ruth I. C. Glasgow
Vivek Singh
Lucía Peña-Pérez
Alissa Wilhalm
Marco F. Moedas
David Moore
Florian A. Rosenberger
Xinping Li
Ilian Atanassov
Mira Saba
Miriam Cipullo
Joanna Rorbach
Anna Wedell
Christoph Freyer
Alexey Amunts
Anna Wredenberg
author_sort Ruth I. C. Glasgow
collection DOAJ
description Abstract S-adenosylmethionine (SAM) is the principal methyl donor in cells and is essential for mitochondrial gene expression, influencing RNA modifications, translation, and ribosome biogenesis. Using direct long-read RNA sequencing in mouse tissues and embryonic fibroblasts, we show that processing of the mitochondrial ribosomal gene cluster fails in the absence of mitochondrial SAM, leading to an accumulation of unprocessed precursors. Proteomic analysis of ribosome fractions revealed these precursors associated with processing and assembly factors, indicating stalled biogenesis. Structural analysis by cryo-electron microscopy demonstrated that SAM-dependent methylation is required for peptidyl transferase centre formation during mitoribosome assembly. Our findings identify a critical role for SAM in coordinating mitoribosomal RNA processing and large subunit maturation, linking cellular methylation potential to mitochondrial translation capacity.
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spelling doaj-art-261cdea9e8a14f608ae2e568edda9cd72025-08-20T03:16:00ZengNature PortfolioNature Communications2041-17232025-06-0116111810.1038/s41467-025-60977-xThe mitochondrial methylation potential gates mitoribosome assemblyRuth I. C. Glasgow0Vivek Singh1Lucía Peña-Pérez2Alissa Wilhalm3Marco F. Moedas4David Moore5Florian A. Rosenberger6Xinping Li7Ilian Atanassov8Mira Saba9Miriam Cipullo10Joanna Rorbach11Anna Wedell12Christoph Freyer13Alexey Amunts14Anna Wredenberg15Department of Medical Biochemistry and Biophysics, Karolinska InstitutetDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetCentre for Inherited Metabolic Diseases, Karolinska University HospitalDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetMax-Planck Institute for Biology of AgeingMax-Planck Institute for Biology of AgeingDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetCentre for Inherited Metabolic Diseases, Karolinska University HospitalDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetUniversity of MünsterDepartment of Medical Biochemistry and Biophysics, Karolinska InstitutetAbstract S-adenosylmethionine (SAM) is the principal methyl donor in cells and is essential for mitochondrial gene expression, influencing RNA modifications, translation, and ribosome biogenesis. Using direct long-read RNA sequencing in mouse tissues and embryonic fibroblasts, we show that processing of the mitochondrial ribosomal gene cluster fails in the absence of mitochondrial SAM, leading to an accumulation of unprocessed precursors. Proteomic analysis of ribosome fractions revealed these precursors associated with processing and assembly factors, indicating stalled biogenesis. Structural analysis by cryo-electron microscopy demonstrated that SAM-dependent methylation is required for peptidyl transferase centre formation during mitoribosome assembly. Our findings identify a critical role for SAM in coordinating mitoribosomal RNA processing and large subunit maturation, linking cellular methylation potential to mitochondrial translation capacity.https://doi.org/10.1038/s41467-025-60977-x
spellingShingle Ruth I. C. Glasgow
Vivek Singh
Lucía Peña-Pérez
Alissa Wilhalm
Marco F. Moedas
David Moore
Florian A. Rosenberger
Xinping Li
Ilian Atanassov
Mira Saba
Miriam Cipullo
Joanna Rorbach
Anna Wedell
Christoph Freyer
Alexey Amunts
Anna Wredenberg
The mitochondrial methylation potential gates mitoribosome assembly
Nature Communications
title The mitochondrial methylation potential gates mitoribosome assembly
title_full The mitochondrial methylation potential gates mitoribosome assembly
title_fullStr The mitochondrial methylation potential gates mitoribosome assembly
title_full_unstemmed The mitochondrial methylation potential gates mitoribosome assembly
title_short The mitochondrial methylation potential gates mitoribosome assembly
title_sort mitochondrial methylation potential gates mitoribosome assembly
url https://doi.org/10.1038/s41467-025-60977-x
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