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J. Biol. Chem., Vol. 280, Issue 4, 2429-2438, January 28, 2005

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RNA Interference Analyses Suggest a Transcript-specific Regulatory Role for Mitochondrial RNA-binding Proteins MRP1 and MRP2 in RNA Editing and Other RNA Processing in Trypanosoma brucei^*

Eva Vondruková, Janny van den Burg¶, Alena Zíková, Nancy Lewis Ernst||, Kenneth Stuart||, Rob Benne¶, and Julius Luke**

From the Institute of Parasitology, Czech Academy of Sciences, and Faculty of Biology, University of South Bohemia, 37005 eské Budjovice, Czech Republic, the ^¶Department of Biochemistry, Academic Medical Center, University of Amsterdam, 1105 AZ Amsterdam, The Netherlands, and the ^||Seattle Biomedical Research Institute and Department of Pathobiology, University of Washington, Seattle, Washington 98195

Mitochondrial RNA-binding proteins MRP1 and MRP2 occur in a heteromeric complex that appears to play a role in U-insertion/deletion editing in trypanosomes. Reduction in the levels of MRP1 (gBP21) and/or MRP2 (gBP25) mRNA by RNA interference in procyclic Trypanosoma brucei resulted in severe growth inhibition. It also resulted in the loss of both proteins, even when only one of the MRP mRNAs was reduced, indicating a mutual dependence for stability. Elimination of the MRPs gave rise to substantially reduced levels of edited CyB and RPS12 mRNAs but little or no reduction of the level of edited Cox2, Cox3, and A6 mRNAs as measured by poisoned primer extension analyses. In contrast, edited NADH-dehydrogenase (ND) subunit 7 mRNA was increased 5-fold in MRP1+2 double knock-down cells. Furthermore, MRP elimination resulted in reduced levels of Cox1, ND4, and ND5 mRNAs, which are never edited, whereas mitoribosomal 12 S rRNA levels were not affected. These data indicate that MRP1 and MRP2 are not essential for RNA editing per se but, rather, play a regulatory role in the editing of specific transcripts and other RNA processing activities.

Received for publication, May 27, 2004 , and in revised form, August 19, 2004.

^* This work was supported by grants from the Grant Agency of the Czech Academy of Sciences (IAA5022302) and the Ministry of Education of the Czech Republic (MMT-123100003) (to J. L.) and by National Institutes of Health Grant AI14102 (to K. S.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Recipient of a European Science Foundation Exchange grant.

^** To whom correspondence should be addressed: Inst. of Parasitology, Braniovská 31, 37005 Ceské Budjovice, Czech Republic. Tel.: 00420-38-7775416; Fax: 00420-38-5310388; E-mail: jula{at}paru.cas.cz.

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