Mitochondrial Polyadenylation Is a One-Step Process Required for mRNA Integrity and tRNA Maturation

PLoS Genet. 2016 May 13;12(5):e1006028. doi: 10.1371/journal.pgen.1006028. eCollection 2016 May.

Abstract

Polyadenylation has well characterised roles in RNA turnover and translation in a variety of biological systems. While polyadenylation on mitochondrial transcripts has been suggested to be a two-step process required to complete translational stop codons, its involvement in mitochondrial RNA turnover is less well understood. We studied knockdown and knockout models of the mitochondrial poly(A) polymerase (MTPAP) in Drosophila melanogaster and demonstrate that polyadenylation of mitochondrial mRNAs is exclusively performed by MTPAP. Further, our results show that mitochondrial polyadenylation does not regulate mRNA stability but protects the 3' terminal integrity, and that despite a lack of functioning 3' ends, these trimmed transcripts are translated, suggesting that polyadenylation is not required for mitochondrial translation. Additionally, loss of MTPAP leads to reduced steady-state levels and disturbed maturation of tRNACys, indicating that polyadenylation in mitochondria might be important for the stability and maturation of specific tRNAs.

MeSH terms

  • Animals
  • Codon, Terminator
  • Drosophila melanogaster / genetics*
  • Gene Knockdown Techniques
  • Mitochondria / genetics
  • Mitochondrial Proteins / biosynthesis
  • Mitochondrial Proteins / genetics
  • Polyadenylation / genetics*
  • Protein Biosynthesis / genetics*
  • RNA, Messenger / genetics*
  • RNA, Mitochondrial
  • RNA, Transfer / genetics

Substances

  • Codon, Terminator
  • Mitochondrial Proteins
  • RNA, Messenger
  • RNA, Mitochondrial
  • mitochondrial messenger RNA
  • RNA, Transfer

Grants and funding

This work was supported by The Swedish Research Council [AWr (VR521-2012-2571)]; Karolinska Institutet [AWr (2013fobi38557), CF (2013fobi37932)]; Åke Wiberg Foundation [AWr (738762088), CF (367990950)]; Stockholm County Council [AWr (K0176-2012)]; Swedish Foundation for Strategic Research [AWr (ICA 12-0017)]; Knut & Alice Wallenberg Foundation [AWr (KAW 20130026)]; AWr is a Ragnar Söderberg fellow (M77/13). The Live Cell Imaging unit is supported by grants from the Knut and Alice Wallenberg Foundation (KAW 2006.0265), the Swedish Research Council (VR213223 and VR213243), the Centre for Biosciences and the Jonasson donation to the School of Technology and Health, Kungliga Tekniska Högskolan, Huddinge, Sweden. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.