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Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation

Pentatricopeptide repeat (PPR) proteins are a large family of proteins that act primarily at different posttranscriptional steps of organellar gene expression. We have previously found that the Schizosaccharomyces pombe PPR protein mpal10 interacts with mitochondrial translational activator Mpa1, an...

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Autores principales: Luo, Ying, Wang, Yirong, Huang, Ying
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8258696/
https://www.ncbi.nlm.nih.gov/pubmed/34119521
http://dx.doi.org/10.1016/j.jbc.2021.100869
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author Luo, Ying
Wang, Yirong
Huang, Ying
author_facet Luo, Ying
Wang, Yirong
Huang, Ying
author_sort Luo, Ying
collection PubMed
description Pentatricopeptide repeat (PPR) proteins are a large family of proteins that act primarily at different posttranscriptional steps of organellar gene expression. We have previously found that the Schizosaccharomyces pombe PPR protein mpal10 interacts with mitochondrial translational activator Mpa1, and both are essential for mitochondrial protein synthesis. However, it is unclear how these two proteins function in mitochondrial protein synthesis in S. pombe. In this study, we further investigated the role of Ppr10 and Mpa1 in mitochondrial protein synthesis. Mitochondrial translational initiation requires two initiation factors, Mti2 and Mti3, which bind to the small subunit of the mitochondrial ribosome (mt-SSU) during the formation of the mitochondrial translational initiation complex. Using sucrose gradient sedimentation analysis, we found that disruption of ppr10, mpa1, or the PPR motifs in Ppr10 impairs the association of Mti2 and Mti3 with the mt-SSU, suggesting that both Ppr10 and Mpa1 may be required for the interaction of Mti2 and Mti3 with the mt-SSU during the assembly of mitochondrial translational initiation complex. Loss of Ppr10 perturbs the association of mitochondrially encoded cytochrome b (cob1) and cytochrome c oxidase subunit 1 (cox1) mRNAs with assembled mitochondrial ribosomes. Proteomic analysis revealed that a fraction of Ppr10 and Mpa1 copurified with a subset of mitoribosomal proteins. The PPR motifs of Ppr10 are necessary for its interaction with Mpa1 and that disruption of these PPR motifs impairs mitochondrial protein synthesis. Our results suggest that Ppr10 and Mpa1 function together to mediate mitochondrial translational initiation.
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spelling pubmed-82586962021-07-12 Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation Luo, Ying Wang, Yirong Huang, Ying J Biol Chem Research Article Pentatricopeptide repeat (PPR) proteins are a large family of proteins that act primarily at different posttranscriptional steps of organellar gene expression. We have previously found that the Schizosaccharomyces pombe PPR protein mpal10 interacts with mitochondrial translational activator Mpa1, and both are essential for mitochondrial protein synthesis. However, it is unclear how these two proteins function in mitochondrial protein synthesis in S. pombe. In this study, we further investigated the role of Ppr10 and Mpa1 in mitochondrial protein synthesis. Mitochondrial translational initiation requires two initiation factors, Mti2 and Mti3, which bind to the small subunit of the mitochondrial ribosome (mt-SSU) during the formation of the mitochondrial translational initiation complex. Using sucrose gradient sedimentation analysis, we found that disruption of ppr10, mpa1, or the PPR motifs in Ppr10 impairs the association of Mti2 and Mti3 with the mt-SSU, suggesting that both Ppr10 and Mpa1 may be required for the interaction of Mti2 and Mti3 with the mt-SSU during the assembly of mitochondrial translational initiation complex. Loss of Ppr10 perturbs the association of mitochondrially encoded cytochrome b (cob1) and cytochrome c oxidase subunit 1 (cox1) mRNAs with assembled mitochondrial ribosomes. Proteomic analysis revealed that a fraction of Ppr10 and Mpa1 copurified with a subset of mitoribosomal proteins. The PPR motifs of Ppr10 are necessary for its interaction with Mpa1 and that disruption of these PPR motifs impairs mitochondrial protein synthesis. Our results suggest that Ppr10 and Mpa1 function together to mediate mitochondrial translational initiation. American Society for Biochemistry and Molecular Biology 2021-06-10 /pmc/articles/PMC8258696/ /pubmed/34119521 http://dx.doi.org/10.1016/j.jbc.2021.100869 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Research Article
Luo, Ying
Wang, Yirong
Huang, Ying
Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation
title Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation
title_full Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation
title_fullStr Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation
title_full_unstemmed Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation
title_short Schizosaccharomyces pombe Ppr10 and Mpa1 together mediate mitochondrial translational initiation
title_sort schizosaccharomyces pombe ppr10 and mpa1 together mediate mitochondrial translational initiation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8258696/
https://www.ncbi.nlm.nih.gov/pubmed/34119521
http://dx.doi.org/10.1016/j.jbc.2021.100869
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