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In vitro guide RNA/mRNA chimaera formation in Trypanosoma brucei RNA editing.

Koslowsky, D. J. ; Göringer, H. Ulrich ; Morales, T. H. ; Stuart, K. :
In vitro guide RNA/mRNA chimaera formation in Trypanosoma brucei RNA editing.
In: Nature, 356 (6372) pp. 807-9. ISSN 0028-0836
[Artikel], (1992)

Kurzbeschreibung (Abstract)

The post-transcriptional processing of various mitochondrial transcripts in kinetoplastids, kRNA editing, adds and removes uridines, producing mature messenger RNAs. This editing seems to be directed by 'guide' RNAs (gRNAs) which are complementary to portions of the mature message. The editing mechanism has been proposed to entail transesterification. Detection of chimaeric gRNA-mRNA molecules, intermediates predicted by transesterification, support this model. We report here the in vitro formation of such chimaeras where endogenous gRNAs are covalently linked to added synthetic mRNA. Addition of gel-purified gRNAs to the standard reaction mix increases chimaera formation. This increase is not observed when the gRNA 3'-hydroxyl group is chemically modified, identifying this terminal hydroxyl as the reactive group. These results provide the first experimental evidence for an in vitro RNA editing event and support the involvement of transesterification as a chemical mechanism.

Typ des Eintrags: Artikel
Erschienen: 1992
Autor(en): Koslowsky, D. J. ; Göringer, H. Ulrich ; Morales, T. H. ; Stuart, K.
Titel: In vitro guide RNA/mRNA chimaera formation in Trypanosoma brucei RNA editing.
Sprache: Englisch
Kurzbeschreibung (Abstract):

The post-transcriptional processing of various mitochondrial transcripts in kinetoplastids, kRNA editing, adds and removes uridines, producing mature messenger RNAs. This editing seems to be directed by 'guide' RNAs (gRNAs) which are complementary to portions of the mature message. The editing mechanism has been proposed to entail transesterification. Detection of chimaeric gRNA-mRNA molecules, intermediates predicted by transesterification, support this model. We report here the in vitro formation of such chimaeras where endogenous gRNAs are covalently linked to added synthetic mRNA. Addition of gel-purified gRNAs to the standard reaction mix increases chimaera formation. This increase is not observed when the gRNA 3'-hydroxyl group is chemically modified, identifying this terminal hydroxyl as the reactive group. These results provide the first experimental evidence for an in vitro RNA editing event and support the involvement of transesterification as a chemical mechanism.

Titel der Zeitschrift, Zeitung oder Schriftenreihe: Nature
Band: 356
(Heft-)Nummer: 6372
Fachbereich(e)/-gebiet(e): Fachbereich Biologie, Biology > Genregulation und RNA-Therapeutika, Postranscriptional Gene Regulation and RNA Therapeutics
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Fachbereich Biologie, Biology
Hinterlegungsdatum: 03 Nov 2011 13:51
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