Chloroplasts are a major destination of protein traffic within leaf cells. Protein import into chloroplasts is mediated by a set of translocon complexes at the chloroplast envelope. Current data indicate that the expression of translocon genes is regulated in a tissue-specific manner, possibly to accommodate the higher import demand of chloroplasts in leaves and the lower demand of plastids in other tissues. We have designed a transgene-based positive screen to isolate mutants disrupted in protein import into plastids. The first locus we isolated, CIA2 , encodes a protein containing a motif conserved within the CCT family of transcription factors. Biochemical analysis indicates that CIA2 is responsible for specific upregulation of the translocon genes atToc33 and atToc75 in leaves. Identification of CIA2 provides new insights into the tissue-specific regulation of translocon gene expression. INTRODUCTIONMost proteins in chloroplasts are encoded by the nucleus and imported post-translationally into chloroplasts. Except for some outer envelope membrane proteins, nucleusencoded chloroplast proteins are synthesized as higher molecular weight precursors with N-terminal extensions called transit peptides. Transit peptides are necessary and sufficient for the import of precursor proteins into chloroplasts. Transport across the double membrane envelope is mediated by a set of translocon components located in the envelope. Several translocon components have been identified from pea chloroplasts by cross-linking or coimmunoprecipitating with importing precursor proteins (reviewed by Schleiff and Soll, 2000). They are collectively named Toc (for translocon at the outer envelope membrane of chloroplasts) and Tic (for translocon at the inner envelope membrane of chloroplasts) proteins (Schnell et al., 1997).Among the Toc components identified, Toc159 is proposed to function as the transit peptide receptor (Perry and Keegstra, 1994;Ma et al., 1996). A considerable amount of evidence indicates that Toc75 is the major component of the protein-translocating channel in the outer membrane (Hinnah et al., 1997;Reumann et al., 1999). The function of Toc34 is not clear. It has been shown to be tightly associated with Toc75 (Seedorf et al., 1995). Arabidopsis has two Toc34 orthologs, atToc34 and atToc33. These two proteins seem to have distinct but overlapping functions (Gutensohn et al., 2000).Techniques such as coimmunoprecipitation and crosslinking, when used to identify translocon components, usually identify abundant and stably associated components. Regulatory components that are present in minute amounts or only transiently, and upstream regulators present in different locations, usually are missed by these techniques.More recently, genetics has been used to study protein import into chloroplasts. Arabidopsis mutants have been found for two translocon components, atToc159 (Bauer et al., 2000) and atToc33 (Jarvis et al., 1998). These mutants confirmed that the translocon components identified by cell biology techniques functi...

Chloroplasts are a major destination of protein traffic within leaf cells. Protein import into chloroplasts is mediated by a set of translocon complexes at the chloroplast envelope. Current data indicate that the expression of translocon genes is regulated in a tissue-specific manner, possibly to accommodate the higher import demand of chloroplasts in leaves and the lower demand of plastids in other tissues. We have designed a transgene-based positive screen to isolate mutants disrupted in protein import into plastids. The first locus we isolated, CIA2 , encodes a protein containing a motif conserved within the CCT family of transcription factors. Biochemical analysis indicates that CIA2 is responsible for specific upregulation of the translocon genes atToc33 and atToc75 in leaves. Identification of CIA2 provides new insights into the tissue-specific regulation of translocon gene expression.