Translational regulation of chloroplast genes - Proteins binding to the 5'-untranslated regions of chloroplast mRNAs in Chlamydomonas reinhardtii

We have examined the effects of illumination, carbon source, and levels of chloroplast protein synthesis on trans-acting proteins that bind to the leaders of five representative chloroplast mRNAs, The accumulation of these five chloroplast mRNAs and the proteins they en code were measured in cells grown under identical conditions, Extracts from all cell types examined contain a minimum set of six chloroplast 5'-untranslated region (UTR)-binding proteins (81, 62, 56, 47, 38, and 15 kDa), Fractionation results suggest that multiple forms of the 81-, 62-, and 47 kDa proteins may exist, A 36-kDa protein was found in all cells except those deficient in chloroplast protein synthesis, Binding of the 81-, 47, and 38-kDa proteins to the rps12 leader is effectively competed by the atpB or rbcL 5'-UTRs, indicating that the same proteins bind to all three leaders. In contrast, these three proteins do not bind to the nuclear encoded alpha-1 tubulin leader, which bound novel proteins of 110, 70, and 43 kDa. Cis-acting sequences within the 5'-UTRs of two chloroplast mRNAs (rps7 and atpB) have been identified which are protected from digestion by RNase T1 by extracts enriched for the 81-, 47-, and 38-kDa proteins.