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The chloroplastNAD(P)H dehydrogenase (NDH )C (ndhC ) andndhK genes partially overlap and are cotranscribed in many plants. We previously reported that the tobaccondhC/K genes are translationally coupled but produce NdhC and NdhK, subunits of the NDH complex, in similar amounts. Generally, translation of the downstream cistron in overlapping mRNAs is very low. Hence, these findings suggested that thendhK cistron is translated not only from thendhC 5′UTR but also by an additional pathway. Using an in vitro translation system from tobacco chloroplasts, we report here that free ribosomes enter, with formylmethionyl-tRNAfMet , at an internal AUG start codon that is located in frame in the middle of the upstreamndhC cistron, translate the 3′ half of thendhC cistron, reach thendhK start codon, and that, at that point, some ribosomes resumendhK translation. We detected a peptide corresponding to a 57-amino-acid product encoded by the sequence from the internal AUG to thendhC stop codon. We propose a model in which the internal initiation site AUG is not designed for synthesizing a functional isoform but for delivering additional ribosomes to thendhK cistron to produce NdhK in the amount required for the assembly of the NDH complex. This pathway is a unique type of translation to produce protein in the needed amount with the cost of peptide synthesis.

Additional pathway to translate the downstream ndhK cistron in partially overlapping ndhC-ndhK mRNAs in chloroplasts