Chloroplast biogenesis: towards the role of localized translation in Arabidopsis
Résumé
Chloroplasts are a major component of plant cells. Until recently, all nuclear-encoded proteins destined to chloroplast were believed to possess an N-terminal and cleavable chloroplast targeting peptide, and to engage the TOC/TIC machinery. However, recent studies have revealed that alternative routes also exists and identified a series of nuclear-encoded proteins imported via such pathways. Recent proteomic studies, conducted by our team (Bouchnak et al., Mol Cell Proteomics 2019), identified a list of cyto-ribosomal subunits associated to chloroplasts, thus suggesting that localized translation might occur at the chloroplast surface. We were recently able to isolate plastid-associated cyto-ribosomes and to decipher their composition when compared to purified whole cell cyto-ribosomes. Interestingly, these plastid-associated cyto-ribosomes contain a few non cyto-ribosomal proteins which might participate to the control of localized translation at the chloroplast surface. These non cyto-ribosomal proteins were first fused to GFP to analyze their subcellular location. Then, we isolated Arabidopsis knock-out mutants affected in the expression of two of these specific proteins and initiated their phenotypical characterization. Finally, with the aim to identify the nature of the nuclear-encoded mRNAs that are translated by these chloroplast-associated cyto-ribosomes, the identification of mRNAs trapped within these plastid-associated cyto-ribosomes was performed.