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Molecular and Cellular Biology, January 2007, p. 777-787, Vol. 27, No. 2
0270-7306/07/$08.00+0 doi:10.1128/MCB.01460-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.
and
Barbara Sollner-Webb*
Department of Biological Chemistry, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205
Received 7 August 2006/ Returned for modification 19 September 2006/ Accepted 27 October 2006
In trypanosome RNA editing, uridylate (U) residues are inserted and deleted at numerous sites within mitochondrial pre-mRNAs by an
20S protein complex that catalyzes cycles of cleavage, U addition/U removal, and ligation. We used RNA interference to deplete TbMP18 (band VII), the last unexamined major protein of our purified editing complex, showing it is essential. TbMP18 is critical for the U-deletional and U-insertional cleavages and for integrity of the
20S editing complex, whose other major components, TbMP99, TbMP81, TbMP63, TbMP52, TbMP48, TbMP42 (bands I through VI), and TbMP57, instead sediment as
10S associations. Additionally, TbMP18 augments editing substrate recognition by the TbMP57 terminal U transferase, possibly aiding the recognition component, TbMP81. The other editing activities and their coordination in precleaved editing remain active in the absence of TbMP18. These data are reminiscent of the data on editing subcomplexes reported by A. Schnaufer et al. (Mol. Cell 12:307-319, 2003) and suggest that these subcomplexes are held together in the
20S complex by TbMP18, as was proposed previously. Our data additionally imply that the proteins are less long-lived in these subcomplexes than they are when held in the complete editing complex. The editing endonucleolytic cleavages being lost when the editing complex becomes fragmented, as upon TbMP18 depletion, should be advantageous to the trypanosome, minimizing broken mRNAs.
Published ahead of print on 13 November 2006.
Present address: Center for Molecular and Mitochondrial Medicine and Genetics, University of California, Irvine, CA.
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