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J. Biol. Chem., Vol. 281, Issue 49, 37661-37667, December 8, 2006

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A Pentatricopeptide Repeat Protein Is a Site Recognition Factor in Chloroplast RNA Editing^*

Kenji Okuda, Takahiro Nakamura, Mamoru Sugita, Toshiyuki Shimizu^¶, and Toshiharu Shikanai¹

From the Graduate School of Agriculture, Kyushu University, 6-10-1 Hakozaki, Higashiku, Fukuoka 812-8581, Japan, the Center for Gene Research, Nagoya University, Chikusa-ku, Nagoya 464-8602, Japan, and the ^¶International Graduate School of Arts and Sciences, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama 230-0045, Japan

In higher plants, RNA editing is a post-transcriptional process that converts C to U in organelle mRNAs. We have previously shown that an Arabidopsis thaliana crr4 mutant is defective with respect to RNA editing for creating the translational initial codon of the plastid ndhD gene (the ndhD-1 site). CRR4 contains 11 pentatricopeptide repeat motifs but does not contain any domains that are likely to be involved in the editing activity. The green fluorescent protein fused to the putative transit peptide of CRR4 targeted the plastid. The recombinant CRR4 expressed in Escherichia coli specifically bound to the 25 nucleotides of the upstream and the 10 nucleotides of the downstream sequences surrounding the editing site of ndhD-1. The target C nucleotide of this editing is not essential for the binding of CRR4. Taken together with the genetic evidence, we conclude that the pentatricopeptide repeat protein CRR4 is a sequence-specific RNA-binding protein that acts as a site recognition factor in plastid RNA editing.

Received for publication, August 25, 2006 , and in revised form, September 28, 2006.

^* This work was supported by Grants-in-aid for Scientific Research on Priority Areas (16085206) and for Creative Scientific Research (17GS0316) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan. 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.

This article was selected as a Paper of the Week.

¹ To whom correspondence should be addressed. Tel. and Fax: 81-92-642-2882; E-mail: shikanai{at}agr.kyushu-u.ac.jp.

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