Description du projet
La logistique des chloroplastes et le rôle des complexes d’importation de protéines
Les cellules sont des usines complexes disposant d’organites individuels liés aux membranes qui contribuent à compartimenter les tâches et les ressources nécessaires à leur réalisation, ce qui améliore l’efficacité. La logistique cellulaire peut s’avérer relativement compliquée dans la mesure où les protéines sont codées dans le noyau, assemblées sur les ribosomes et acheminées vers leur lieu de travail à travers la cellule. Les chloroplastes sont les organites photosynthétiques des cellules végétales, et ils disposent de machines moléculaires spécialisées qui assurent la translocation des protéines à travers leur enveloppe membranaire à double paroi. L’appareil dit translocon, se trouvant au niveau de la membrane d’enveloppe extérieure des chloroplastes (TOC), possède au moins deux configurations principales correspondant à la charge de pré-protéines importées. Le projet TOC-maker, financé par l’UE, étudie les détails du processus d’assemblage et l’organisation structurelle finale des complexes TOC.
Objectif
Plants convert solar energy into chemical energy by the process called photosynthesis in a specialized compartment of the cell known as chloroplasts. Chloroplasts are majorly enriched with nucleus-encoded proteins and to import them, chloroplast outer and inner envelope membranes are equipped with apparatus called the TOC and TIC translocons, respectively. For the TOC apparatus, there are two major configurations, TOC-P and TOC-H, reported so far, which import highly abundant, Photosynthetic and Housekeeping pre-proteins, respectively. TOC-P and TOC-H are multiprotein complexes which must be specifically assembled for proper development and homeostasis of the chloroplast. Due to the dynamic nature of the translocons, component synthesis and assembly must be rapid and tightly coupled, making the process difficult to investigate. Thus, understanding the mechanisms of the assembly process is both challenging and exciting. Biogenesis of TOC complexes is rapidly enhanced during chloroplast development or de-etiolation, and I will exploit this process to investigate the assembly of different TOC configurations, using the model plant Arabidopsis thaliana. For this purpose, transgenic plants expressing epitope-tagged TOC components and cells expressing nascent polypeptides of TOC components with stalled ribosomes will be generated. Proteins transiently interacting with new TOC components, which are predicted to assist integration and assembly of the TOC complex, will be studied by using affinity purification, pulse-chase experiments, and other biochemical techniques, and thus a sequence of assembly events will be elucidated. Although the molecular composition of the TOC protein import machinery has been well studied, the detailed structural organization of TOC complexes has not yet been elucidated. I will address this knowledge gap by analysing affinity-purified TOC complexes from mature chloroplasts at high resolution by cryo-electron microscopy.
Champ scientifique
- engineering and technologyenvironmental engineeringenergy and fuelsrenewable energysolar energy
- natural sciencesbiological sciencesbiochemistrybiomoleculesproteins
- natural sciencesphysical sciencesopticsmicroscopy
- medical and health sciencesbasic medicinephysiologyhomeostasis
- natural sciencesbiological sciencesbotany
Mots‑clés
Programme(s)
Régime de financement
MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)Coordinateur
OX1 2JD Oxford
Royaume-Uni