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Molecular and Cellular Biology, January 2008, p. 122-130, Vol. 28, No. 1
0270-7306/08/$08.00+0     doi:10.1128/MCB.01374-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

RNA Editing in Trypanosoma brucei Requires Three Different Editosomes{triangledown} ,{dagger}

Jason Carnes,1,2 James Raffaello Trotter,1,{ddagger} Adam Peltan,3 Michele Fleck,1 and Kenneth Stuart1,2*

Seattle Biomedical Research Institute, Seattle, Washington 98109,1 Department of Pathobiology, University of Washington, Seattle, Washington 98195,2 Immunology and Infection Unit, Department of Biology, University of York, Heslington, York YO10 5YW, United Kingdom3

Received 31 July 2007/ Returned for modification 20 September 2007/ Accepted 8 October 2007

Trypanosoma brucei has three distinct ~20S editosomes that catalyze RNA editing by the insertion and deletion of uridylates. Editosomes with the KREN1 or KREN2 RNase III type endonucleases specifically cleave deletion and insertion editing site substrates, respectively. We report here that editosomes with KREPB2, which also has an RNase III motif, specifically cleave cytochrome oxidase II (COII) pre-mRNA insertion editing site substrates in vitro. Conditional repression and mutation studies also show that KREPB2 is an editing endonuclease specifically required for COII mRNA editing in vivo. Furthermore, KREPB2 expression is essential for the growth and survival of bloodstream forms. Thus, editing in T. brucei requires at least three compositionally and functionally distinct ~20S editosomes, two of which distinguish between different insertion editing sites. This unexpected finding reveals an additional level of complexity in the RNA editing process and suggests a mechanism for how the selection of sites for editing in vivo is controlled.

* Corresponding author. Mailing address: Seattle Biomedical Research Institute, 307 Westlake Ave N, Suite 500, Seattle, WA 98109. Phone: (206) 256-7316. Fax: (206) 256-7229. E-mail: kstuart{at}u.washington.edu

{triangledown} Published ahead of print on 22 October 2007.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} Present address: Centre for Molecular Microbiology and Infection, Division of Cell and Molecular Biology, Imperial College London, South Kensington Campus, London SW7 2AZ, United Kingdom.

Molecular and Cellular Biology, January 2008, p. 122-130, Vol. 28, No. 1
0270-7306/08/$08.00+0     doi:10.1128/MCB.01374-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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