Biochemical methods for analysis of kinetoplastid RNA editing

Methods. 1998 May;15(1):15-26. doi: 10.1006/meth.1998.0602.

Abstract

RNA editing is a posttranscriptional process involving mRNAs [reviewed by K. Stuart et al. (1997) Microbiol. Mol. Biol. Rev. 61, 105-120; G. J. Arts and R. Benne (1996) Biochim. Biophys. Acta 1307, 39-54; and S. L. Hajduk and R. S. Sabatini (1996) in Molecular Biology of Parasitic Protozoa (Smith, D. S., and Parsons, M., Eds.), pp. 134-158, Oxford Univ. Press, Oxford] and tRNAs [K. M. Lonergan and M. Gray (1993) Science 259, 812-816] that has now been described in an increasing number of eukaryotic organisms. In this process sequences differ from their gene sequences by the addition, removal, or conversion of specific ribonucleotides. RNA editing was first described within the mitochondrion of kinetoplastid protozoa. Several of the mitochondrial mRNAs in these flagellates have uridine residues inserted and deleted at specific sites. In some cases, more than 50% of the mRNA is created by RNA editing. In this article, we describe some of the biochemical methods used in analyzing the process of RNA editing in kinetoplastid mitochondria.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Fractionation
  • DNA, Kinetoplast
  • Dimerization
  • Endonucleases / metabolism
  • Genetic Techniques
  • Ligases / metabolism
  • Mitochondria
  • RNA Editing*
  • RNA Helicases
  • RNA Nucleotidyltransferases / metabolism
  • RNA, Protozoan / genetics*
  • RNA, Protozoan / metabolism
  • Ribonucleoproteins / metabolism
  • Transcription, Genetic
  • Trypanosoma brucei brucei / genetics*
  • Trypanosoma brucei brucei / growth & development
  • Trypanosoma brucei brucei / ultrastructure
  • Uridine Triphosphate / metabolism

Substances

  • DNA, Kinetoplast
  • RNA, Protozoan
  • Ribonucleoproteins
  • RNA Nucleotidyltransferases
  • UTP-RNA uridylyltransferase
  • Endonucleases
  • RNA Helicases
  • Ligases
  • Uridine Triphosphate