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J. Biol. Chem., Vol. 283, Issue 12, 7314-7319, March 21, 2008

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Cross-competition in Editing of Chloroplast RNA Transcripts in Vitro Implicates Sharing of Trans-factors between Different C Targets^*

From the Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853

CU plant organellar RNA editing is required for the translation of evolutionarily conserved and functional proteins. 28 different C targets of RNA editing have been identified in maize chloroplasts, and hundreds of Cs are edited in mitochondria. Mutant analysis in Arabidopsis has indicated that absence of a single site-specific recognition protein can result in loss of editing of a single C target, raising the possibility that each C target requires a recognition protein. Here we show that transcripts encompassing two editing sites, ZMrpoB C467 and ZMrps14 C80, can compete editing activity from each other in vitro despite limited sequence similarity. The signal causing competition overlaps a 5'-cis element required for editing efficiency. A single five-nucleotide mutation spanning the region from –20 to –16 relative to the edited C of rpoB C467 is sufficient to eliminate its substrate editing as well as its ability to compete editing activity from rps14 C80 substrates. A corresponding mutation in an rps14 C80 competitor likewise eliminated its ability to compete editing activity from rpoB C467 substrates. Taken together, our results indicate that the RNA sequences mediating both editing efficiency and cross-competition are highly similar and that a common protein is involved in their editing. Sharing of trans-factors can facilitate editing of the large number of different C targets in plant organelles so that a different protein factor would not be required for every editing site.

Received for publication, November 26, 2007 , and in revised form, January 3, 2008.

^* This work was supported by National Science Foundation MCB-0344007 and MCB-0716888 (to M. R. H.) and National Institutes of Health graduate traineeships (to W. P. H. and M. L. H.). 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.

The on-line version of this article (available at http://www.jbc.org) contains a supplemental table.

This article was selected as a Paper of the Week.

¹ To whom correspondence should be addressed: Dept. of Molecular Biology and Genetics, Biotech Bldg., Cornell University, Ithaca, NY 14853. Tel.: 607-254-4833; Fax: 607-255-6249; E-mail: mrh5{at}cornell.edu.

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