The pentatricopeptide repeat protein MTSF2 stabilizes a nad1 precursor transcript and defines the 3΄ end of its 5΄-half intron

Nucleic Acids Res. 2017 Jun 2;45(10):6119-6134. doi: 10.1093/nar/gkx162.

Abstract

RNA expression in plant mitochondria implies a large number of post-transcriptional events in which transcript processing and stabilization are essential. In this study, we analyzed the function of the Arabidopsis mitochondrial stability factor 2 gene (MTSF2) and show that the encoded pentatricopeptide repeat protein is essential for the accumulation of stable nad1 mRNA. The production of mature nad1 requires the assembly of three independent RNA precursors via two trans-splicing reactions. Genetic analyses revealed that the lack of nad1 in mtsf2 mutants results from the specific destabilization of the nad1 exons 2-3 precursor transcript. We further demonstrated that MTSF2 binds to its 3΄ extremity with high affinity, suggesting a protective action by blocking exoribonuclease progression. By defining the 3΄ end of nad1 exons 2-3 precursor, MTSF2 concomitantly determines the 3΄ extremity of the first half of the trans-intron found at the end of the transcript. Therefore, binding of the MTSF2 protein to nad1 exons 2-3 precursor evolved both to stabilize the transcript and to define a 3΄ extremity compatible with the trans-splicing reaction needed to reconstitute mature nad1. We thus reveal that the range of transcripts stabilized by association with protective protein on their 3΄ end concerns also mitochondrial precursor transcripts.

Publication types

  • Comparative Study

MeSH terms

  • Amino Acid Sequence
  • Arabidopsis / genetics*
  • Arabidopsis / growth & development
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Arabidopsis Proteins / physiology
  • Base Sequence
  • Binding Sites
  • CRISPR-Cas Systems
  • Electron Transport Complex I / metabolism
  • Exons
  • Introns / genetics
  • Mitochondria / genetics
  • Mitochondria / metabolism*
  • NADH Dehydrogenase / genetics*
  • Plants, Genetically Modified
  • Protein Binding
  • RNA / metabolism*
  • RNA Precursors / metabolism*
  • RNA Splicing
  • RNA Stability
  • RNA, Mitochondrial
  • RNA, Plant / metabolism*
  • RNA-Binding Protein EWS / genetics
  • RNA-Binding Protein EWS / physiology*
  • Recombinant Fusion Proteins / metabolism
  • Sequence Alignment
  • Sequence Homology, Nucleic Acid

Substances

  • Arabidopsis Proteins
  • MTSF2 protein, Arabidopsis
  • RNA Precursors
  • RNA, Mitochondrial
  • RNA, Plant
  • RNA-Binding Protein EWS
  • Recombinant Fusion Proteins
  • RNA
  • NADH Dehydrogenase
  • Nad1 protein, Arabidopsis
  • Electron Transport Complex I