RNA editing in Physarum mitochondria: assays and biochemical approaches.
Mitochondrial RNAs in the myxomycete Physarum polycephalum differ from the templates from which they are transcribed in defined ways. Most transcripts contain nucleotides that are not present in their respective genes. These "extra" nucleotides are added during RNA synthesis by an unknown mechanism. Other differences observed between Physarum mitochondrial RNAs and the mitochondrial genome include nucleotide deletions, C to U changes, and the replacement of one nucleotide for another at the 5' end of tRNAs. All of these alterations are remarkably precise and highly efficient in vivo. Many of these editing events can be replicated in vitro, and here we describe both the in vitro systems used to study editing in Physarum mitochondria and the assays that have been developed to assess the extent of editing of RNAs generated in these systems at individual sites.
Byrne
Elaine M
EM
Trinity Centre for Engineering, Trinity College Dublin, Dublin, Ireland.
Visomirski-Robic
Linda
L
Cheng
Yu-Wei
YW
Rhee
Amy C
AC
Gott
Jonatha M
JM
D000818:Animals; D001671:Biochemistry / Q000379:methods*; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005786:Gene Expression Regulation; D008928:Mitochondria / Q000378:metabolism; D009711:Nucleotides / Q000737:chemistry; D010804:Physarum / Q000235:genetics* / Q000378:metabolism*; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D017393:RNA Editing; D012333:RNA, Messenger / Q000378:metabolism; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism; D014158:Transcription, Genetic
D004277:DNA, Single-Stranded; D009711:Nucleotides; D012333:RNA, Messenger; D012313:RNA; D004262:DNA Restriction Enzymes; D015719:Single-Strand Specific DNA and RNA Endonucleases