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Kinetoplastid RNA editing does not require the terminal 3′ hydroxyl of guide RNA, but modifications to the guide RNA terminus can inhibit in vitro U insertion

Published online by Cambridge University Press:  01 July 1999

MOFFETT L.K. BURGESS ,
STEFAN HEIDMANN  and
KENNETH STUART
MOFFETT L.K. BURGESS
Affiliation:
Seattle Biomedical Research Institute, Seattle, Washington 98109, USA, and Department of Pathobiology, University of Washington, Seattle, Washington 98195, USA
STEFAN HEIDMANN
Affiliation:
Seattle Biomedical Research Institute, Seattle, Washington 98109, USA, and Department of Pathobiology, University of Washington, Seattle, Washington 98195, USA Present address: Department of Genetics, University of Bayreuth, 95440 Bayreuth, Germany.
KENNETH STUART
Affiliation:
Seattle Biomedical Research Institute, Seattle, Washington 98109, USA, and Department of Pathobiology, University of Washington, Seattle, Washington 98195, USA
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Abstract

During RNA editing in kinetoplastid parasites, trans-acting guide RNAs (gRNAs) direct the insertion and deletion of U residues at precise sites in mitochondrial pre-mRNAs. We show here that some modifications to the 3′ terminal ribose of gRNA inhibit its ability to direct in vitro U insertion. However, we found that gRNAs lacking this moiety in some circumstances support in vitro editing. Thus, the 3′ OH is not required. Inhibition resulting from gRNA modification can be overcome by increasing the gRNA–pre-mRNA base-pairing potential upstream of the editing site, suggesting an importance for this interaction to productive processing.

Keywords

guide RNAKinetoplastidaeRNA editingtrypanosomes
Type
Research Article
Information
RNA , Volume 5 , Issue 7 , July 1999 , pp. 883 - 892
Copyright
© 1999 RNA Society

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