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The specificity of nucleotide removal during RNA editing in Trypanosoma brucei

Published online by Cambridge University Press:  11 January 2002

SOBOMABO D. LAWSON ,
ROBERT P. IGO ,
REZA SALAVATI  and
KENNETH D. STUART
SOBOMABO D. LAWSON
Affiliation:
Seattle Biomedical Research Institute and Pathobiology Department, University of Washington, Seattle, Washington 98109-1651, USA
ROBERT P. IGO
Affiliation:
Seattle Biomedical Research Institute and Pathobiology Department, University of Washington, Seattle, Washington 98109-1651, USA
REZA SALAVATI
Affiliation:
Seattle Biomedical Research Institute and Pathobiology Department, University of Washington, Seattle, Washington 98109-1651, USA
KENNETH D. STUART
Affiliation:
Seattle Biomedical Research Institute and Pathobiology Department, University of Washington, Seattle, Washington 98109-1651, USA
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Abstract

RNA editing in Trypanosoma brucei produces mature mRNAs by posttranscriptional insertion and deletion of uridylates (Us) by a series of catalytic steps, which include endoribonucleolytic cleavage, 3′ terminal addition or removal of Us, and RNA ligation. Preedited mRNA (pre-mRNA) and guide RNA (gRNA) that are mutated at or near the editing site (ES) were used to examine the effects on the specificity of in vitro editing. Sequences that are not predicted to form a gRNA/pre-mRNA base pair immediately 5′ to the ES still supported accurate editing. Substitution of a non-U nucleotide at various positions within a stretch of Us that are normally removed from the ES resulted in deletion of only the Us that were 3′ to the substituted nucleotide. Overall, ES selection by the endoribonuclease, the specificity of the 3′ exoribonuclease for Us, and ligation appear to act in concert to ensure the production of accurately edited RNA.

Keywords

3′ exonucleasegRNAmechanismuridylate specificity
Type
Research Article
Information
RNA , Volume 7 , Issue 12 , December 2001 , pp. 1793 - 1802
Copyright
2001 RNA Society

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