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Genes 2016, 7(12), 128; doi:10.3390/genes7120128

Electroporation of DNA into Physarum polycephalum Mitochondria: Effects on Transcription and RNA Editing in Isolated Organelles

1
Center for RNA Molecular Biology, Case Western Reserve University, Cleveland, OH 44120, USA
2
Visual Sciences Research Center, Case Western Reserve University, Cleveland, OH 44120, USA
Current address: Department of Stem Cell Biology and Regenerative Medicine, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
*
Author to whom correspondence should be addressed.
Academic Editor: H. Ulrich Göringer
Received: 31 October 2016 / Revised: 2 December 2016 / Accepted: 5 December 2016 / Published: 14 December 2016
(This article belongs to the Special Issue RNA Editing)
View Full-Text   |   Download PDF [2705 KB, uploaded 14 December 2016]   |  

Abstract

Mitochondrial RNAs in the acellular slime mold Physarum polycephalum contain nucleotides that are not encoded in the mitochondrial genes from which they are transcribed. These site-specific changes are quite extensive, comprising ~4% of the residues within mRNAs and ~2% of rRNAs and tRNAs. These “extra” nucleotides are added co-transcriptionally, but the means by which this is accomplished have not been elucidated. The cox1 mRNA also contains four sites of C to U changes, which occur post-transcriptionally, most likely via targeted deamination. The currently available in vitro systems for studying P. polycephalum editing are limited in that the template is the entire ~63,000 bp mitochondrial genome. This presents a significant challenge when trying to define the signals that specify editing sites. In an attempt to overcome this issue, a method for introducing DNA into isolated P. polycephalum mitochondria via electroporation has been developed. Exogenous DNA is expressed, but the transcripts synthesized from these templates are not edited under the conditions tested. However, transcripts derived from the mitochondrial genome are accurately edited after electroporation, indicating that the editing machinery is still functional. These findings suggest that this method may ultimately provide a feasible approach to elucidating editing signals. View Full-Text
Keywords: RNA editing; Physarum polycephalum; mitochondria; electroporation RNA editing; Physarum polycephalum; mitochondria; electroporation
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MDPI and ACS Style

Gott, J.M.; Naegele, G.M.; Howell, S.J. Electroporation of DNA into Physarum polycephalum Mitochondria: Effects on Transcription and RNA Editing in Isolated Organelles. Genes 2016, 7, 128.

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