Identifying ChloroPlast Ribosome rescue mechanisms under genetically induced and...
Identifying ChloroPlast Ribosome rescue mechanisms under genetically induced and environmental stresses
Photosynthesis produces all organic material on Earth. In plants, photosynthesis depends on active protein synthesis in chloroplasts to set up photosynthetic complexes. Due to their endosymbiotic origin (from cyanobacteria), chlor...
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Descripción del proyecto
Photosynthesis produces all organic material on Earth. In plants, photosynthesis depends on active protein synthesis in chloroplasts to set up photosynthetic complexes. Due to their endosymbiotic origin (from cyanobacteria), chloroplasts contain an independent genome and internal transcription and translation machinery with bacterial type 70S-like ribosomes. Ribosomes may stall due to transcription errors, misprocessed transcripts, translation mistakes, missing tRNA, mutations, or strong mRNA-folding. Ribosome stalling ties up the translation machinery, it is persistent at trace levels but can be exacerbated by environmental stress. Therefore, ribosome rescue mechanisms have evolved in all life domains to resolve stalled ribosomes. Despite its critical role in maintaining photosynthesis and extensive study in eukaryotes and bacteria, little is known about ribosome rescue mechanisms in chloroplasts.
In this project, we will characterise chloroplast rescue factors. First, we will examine ArfB, the only known ribosome rescue mechanism ortholog found in chloroplasts by exposing ArfB knockout mutants to translation-stalling inducing stresses, investigating conserved protein motifs, subcellular localisation and overexpression. Additionally, we will study ribosome-stalling effects on chloroplast translation, photosynthesis and metabolism by genetically modifying the chloroplast translation machinery. Finally, we will directly search for novel ribosome rescue factors by identifying proteins physically engaged with stalled chloroplast ribosomes and evaluating their function.
Elucidating chloroplast ribosome rescue mechanisms will improve our understanding of the complete photosynthesis support apparatus and inform future biotechnology and crop improvement. Following this fellowship I will leverage the knowledge, methods and transferable skills I acquire to study chloroplast genetics, translation and nuclear-chloroplast interactions in my future lab.