Mitochondrial electroporation and in organello RNA editing of chimeric atp6 transcripts

Mol Genet Genomics. 2005 Apr;273(2):130-6. doi: 10.1007/s00438-005-1117-x. Epub 2005 Feb 24.

Abstract

The Sorghum bicolor atp6-1 gene and chimeric atp6 genes with additional maize sequences were introduced into isolated maize mitochondria via electroporation. Transcripts isolated after in vitro incubation of the transformed organelles were then analysed for RNA editing. Transcripts of the S. bicolor atp6-1 gene, and the RNAs obtained from most of chimeric sorghum-maize atp6 gene constructs tested, were not edited. However, the transcript of one engineered chimeric gene comprising the 5'untranslated sequence and a segment of the N-terminal ORF of the maize atp6 combined with the sorghum atp6 core ORF and 3'untranslated sequence was found to be partially edited. We were able to exclude low RNA stability or insufficient editing capacity as the reason for failure to edit in the other instances. Instead, the data indicate that the maize sequence in the edited fusion transcript provides a structural motif or binding site for a transcript-specific editing factor.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Electroporation
  • Genetic Vectors
  • Mitochondria / metabolism*
  • Mitochondrial Proton-Translocating ATPases / genetics*
  • Mitochondrial Proton-Translocating ATPases / metabolism
  • Molecular Sequence Data
  • Oligonucleotides
  • RNA Editing / genetics*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, DNA
  • Sorghum / genetics*
  • Zea mays / genetics*

Substances

  • Oligonucleotides
  • Mitochondrial Proton-Translocating ATPases