Evolution of RNA editing sites in the mitochondrial small subunit rRNA of the Myxomycota

Methods Enzymol. 2007:424:197-220. doi: 10.1016/S0076-6879(07)24009-1.

Abstract

Because of their unique and unprecedented character, it is often difficult to imagine how and why the different, diverse types of RNA editing have evolved. Information about the evolution of a particular RNA editing system can be obtained by comparing RNA editing characteristics in contemporary organisms whose phylogenetic relationships are known so that editing patterns in ancestral organisms can be inferred. This information can then be used to build models of the origins, constraints, variability, and mechanisms of RNA editing. As an example of the types of information that can be obtained from these analyses, we describe how we have used cDNA, covariation, and phylogenetic analyses to study the evolution of the variation in RNA editing site location in the core region of the small subunit rRNA gene in the mtDNA of seven myxomycetes, including Physarum polycephalum. We find that the unique type of insertional RNA editing present in mitochondria of P. polycephalum is also present in the mitochondrial small subunit (SSU) rRNA of the other six myxomycetes. As in Physarum, this editing predominantly consists of cytidine insertions, but also includes uridine insertions and certain dinucleotide insertions such that any of the four canonical ribonucleotides can be inserted. Although the characteristics of RNA editing in these organisms are the same as in Physarum, the location of the insertion sites varies among the seven organisms relative to the conserved primary sequence and secondary structure of the rRNA. Nucleotide insertions have been identified at 29 different sites within this core region of the rRNA, but no one organism has more than 10 of these insertion sites, suggesting that editing sites have been created and/or eliminated since the divergence of these organisms. To determine the order in which editing sites have been created or eliminated, the sequences of the mitochondrial SSU rRNA have been aligned and this alignment has been used to produce phylogenetic trees showing the sequence relationship of these organisms. These phylogenetic trees are congruent with phylogenetic trees predicted by alignment of nuclear rDNA sequences. These trees indicate that editing sites change rapidly relative to mtDNA sequence divergence and suggest that some editing sites have been created more than once during the evolution of the Myxomycota.

MeSH terms

  • Animals
  • Base Sequence
  • Binding Sites
  • Biochemistry / methods*
  • Cloning, Molecular
  • DNA Primers / chemistry
  • Genetic Techniques*
  • Molecular Sequence Data
  • Myxomycetes / genetics*
  • Nucleic Acid Conformation
  • Phylogeny
  • RNA / metabolism
  • RNA Editing*
  • RNA, Mitochondrial
  • RNA, Ribosomal / genetics*
  • Sequence Homology, Nucleic Acid

Substances

  • DNA Primers
  • RNA, Mitochondrial
  • RNA, Ribosomal
  • RNA