RNA editing complex interactions with a site for full-round U deletion in Trypanosoma brucei

Anastasia Sacharidou, Catherine Cifuentes-Rojas, Kari Halbig, Alfredo Hernandez, Lawrence J. Dangott, Monica De Nova-Ocampo, Jorge Cruz-Reyes

Research output: Contribution to journalArticle

11 Scopus citations

Abstract

Trypanosome U insertion and U deletion RNA editing of mitochondrial pre-mRNAs is catalyzed by multisubunit editing complexes as directed by partially complementary guide RNAs. The basic enzymatic activities and protein composition of these high-molecular mass complexes have been under intense study, but their specific protein interactions with functional pre-mRNA/gRNA substrates remains unknown. We show that editing complexes purified through extensive ion-exchange chromatography and immunoprecipitation make specific cross-linking interactions with A6 pre-mRNA containing a single 32P and photoreactive 4-thioU at the scissile bond of a functional site for full-round U deletion. At least four direct protein-RNA contacts are detected at this site by cross-linking. All four interactions are stimulated by unpaired residues just 5′ of the pre-mRNA/ gRNA anchor duplex, but strongly inhibited by pairing of the editing site region. Furthermore, competition analysis with homologous and heterologous transcripts suggests preferential contacts of the editing complex with the mRNA/gRNA duplex substrate. This apparent structural selectivity suggests that the RNA-protein interactions we observe may be involved in recognition of editing sites and/or catalysis in assembled complexes.

Original languageEnglish (US)
Pages (from-to)1219-1228
Number of pages10
JournalRNA
Volume12
Issue number7
DOIs
StatePublished - 2006
Externally publishedYes

Keywords

  • Editing complexes
  • RNA editing
  • RNA-protein interactions
  • Trypanosoma brucei

ASJC Scopus subject areas

  • Molecular Biology

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  • Cite this

    Sacharidou, A., Cifuentes-Rojas, C., Halbig, K., Hernandez, A., Dangott, L. J., De Nova-Ocampo, M., & Cruz-Reyes, J. (2006). RNA editing complex interactions with a site for full-round U deletion in Trypanosoma brucei. RNA, 12(7), 1219-1228. https://doi.org/10.1261/rna.2295706