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J. Biol. Chem., Vol. 279, Issue 9, 7819-7825, February 27, 2004

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The Effect of RNA Interference Down-regulation of RNA Editing 3'-Terminal Uridylyl Transferase (TUTase) 1 on Mitochondrial de Novo Protein Synthesis and Stability of Respiratory Complexes in Trypanosoma brucei^*

Martina Nebohácová, Dmitri A. Maslov, Arnold M. Falick¶, and Larry Simpson**

From the Howard Hughes Medical Institute and Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, California 90095, Department of Biology, University of California, Riverside, California 92521, and ^¶Howard Hughes Medical Institute Mass Spectrometry Laboratory, Department of Molecular and Cell Biology, University of California, Berkeley, California 94720

Inhibition of RNA editing by down-regulation of expression of the mitochondrial RNA editing TUTase 1 by RNA interference had profound effects on kinetoplast biogenesis in Trypanosoma brucei procyclic cells. De novo synthesis of the apocytochrome b and cytochrome oxidase subunit I proteins was no longer detectable after 3 days of RNAi. The effect on protein synthesis correlated with a decline in the levels of the assembled mitochondrial respiratory complexes III and IV, and also cyanide-sensitive oxygen uptake. The steady-state levels of nuclear-encoded subunits of complexes III and IV were also significantly decreased. Because the levels of the corresponding mRNAs were not affected, the observed effect was likely due to an increased turnover of these imported mitochondrial proteins. This induced protein degradation was selective for components of complexes III and IV, because little effect was observed on components of the F₁·F₀-ATPase complex and on several other mitochondrial proteins.

Received for publication, October 15, 2003 , and in revised form, December 7, 2003.

^* This work was supported in part by National Institutes of Health Grant AI09102 (to L. 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.

^** To whom correspondence should be addressed: Howard Hughes Medical Institute, 6780 MacDonald Research Laboratories, 675 Charles Young Dr. S., Los Angeles, CA 90095-1662. Tel.: 310-825-4215; Fax: 310-206-8967; E-mail: simpson{at}kdna.ucla.edu.

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				L. E. Yu and D. J. Koslowsky Interactions of mRNAs and gRNAs involved in trypanosome mitochondrial RNA editing: Structure probing of a gRNA bound to its cognate mRNA RNA, June 1, 2006; 12(6): 1050 - 1060. [Abstract] [Full Text] [PDF]