Trans-splicing and RNA editing of LSU rRNA in Diplonema mitochondria

Nucleic Acids Res. 2014 Feb;42(4):2660-72. doi: 10.1093/nar/gkt1152. Epub 2013 Nov 19.

Abstract

Mitochondrial ribosomal RNAs (rRNAs) often display reduced size and deviant secondary structure, and sometimes are fragmented, as are their corresponding genes. Here we report a mitochondrial large subunit rRNA (mt-LSU rRNA) with unprecedented features. In the protist Diplonema, the rnl gene is split into two pieces (modules 1 and 2, 534- and 352-nt long) that are encoded by distinct mitochondrial chromosomes, yet the rRNA is continuous. To reconstruct the post-transcriptional maturation pathway of this rRNA, we have catalogued transcript intermediates by deep RNA sequencing and RT-PCR. Gene modules are transcribed separately. Subsequently, transcripts are end-processed, the module-1 transcript is polyuridylated and the module-2 transcript is polyadenylated. The two modules are joined via trans-splicing that retains at the junction ∼ 26 uridines, resulting in an extent of insertion RNA editing not observed before in any system. The A-tail of trans-spliced molecules is shorter than that of mono-module 2, and completely absent from mitoribosome-associated mt-LSU rRNA. We also characterize putative antisense transcripts. Antisense-mono-modules corroborate bi-directional transcription of chromosomes. Antisense-mt-LSU rRNA, if functional, has the potential of guiding concomitantly trans-splicing and editing of this rRNA. Together, these findings open a window on the investigation of complex regulatory networks that orchestrate multiple and biochemically diverse post-transcriptional events.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Euglenozoa / genetics*
  • Mitochondria / genetics
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • RNA / chemistry
  • RNA / genetics
  • RNA / metabolism*
  • RNA Editing*
  • RNA, Antisense / analysis
  • RNA, Mitochondrial
  • RNA, Ribosomal / chemistry
  • RNA, Ribosomal / genetics
  • RNA, Ribosomal / metabolism*
  • Ribosome Subunits, Large, Eukaryotic / metabolism
  • Trans-Splicing*

Substances

  • RNA, Antisense
  • RNA, Mitochondrial
  • RNA, Ribosomal
  • RNA

Associated data

  • GENBANK/KF633465
  • GENBANK/KF633466
  • GENBANK/KF633467
  • GENBANK/KF633468