Membranous translation platforms in the chloroplast of Chlamydomonas reinhardtii

A small genome in chloroplasts encodes polypeptide subunits of the photosynthetic electron transport complexes in the membranes of thylakoid vesicles in the chloroplast stroma. Trans-membrane subunits of these complexes undergo co-translational membrane insertion during their synthesis by ribosomes of the bacterial-like genetic system of this semiautonomous organelle. While thylakoid membranes are sites of translation, evidence in the unicellular alga Chlamydomonas reinhardtii supports translation also on non-canonical membranes in a discrete translation-zone in the chloroplast. To characterize the membranous platforms for translation and the biogenesis of thylakoid membrane, we profiled membranes during chloroplast development, using the yellow-in-the-dark 1 mutant, and carried out proteomic analyses on membranes of interest. The results support roles of two membrane types in preliminary and ongoing stages of translation: a "low-density membrane" and a denser "chloroplast translation membrane", respectively. These roles are based on correlations of the relative levels of each membrane type and the translational status of the chloroplast before, during and after chloroplast differentiation and results of proteomic analyses. Our results support a model of photosynthesis complex biogenesis in a spatiotemporal "assembly line" involving LDM and CTM as sequential stages leading to photosynthetic thylakoid membranes.