Université de Montpellier 2 Sciences et Techniques du Languedoc, Montpellier
Abstract
Fe is essential for all cells because it is the cofactor of numerous proteins, however, excess free Fe is potentially deleterious for the cell. Ferritins are multimeric proteins, present in all the kingdoms of life that can store iron in a safe and bioavailable form. In mammals, ferritins are the main Fe store. They have been predicted to fulfil the same function in plants, but direct evidences are lacking. In plants, ferritin synthesis in response to iron overload is mainly regulated at the transcriptional level, whereas, in animals, it is mainly regulated at the post-transcriptional level by the aconitase dependent IRP/IRE system. The aims of my PhD project were: (i) to elucidate ferritin function in plant physiology and (ii) to decipher the signaling pathway leading to ferritin accumulation in response to iron overload. (i) To directly study ferritin function in plants, a loss-of-function approach was developed in Arabidopsis. We present evidence that ferritins do not constitute the major iron pool either in seeds for seedling development or in leaves for proper functioning of the photosynthetic apparatus. The loss of ferritins in vegetative and reproductive organs resulted in sensitivity to excess iron. Furthermore, the absence of ferritin led to a strong deregulation of expression of several metal transporter genes in the stalk, over-accumulation of iron in reproductive organs, and a decrease in fertility. Finally, I showed that in the absence of ferritin, plants had higher levels of ROS, and increased activity of enzymes involved in their detoxification. Ferritins are also involved in iron-detoxification during senescence to avoid ROS accumulation. Seeds ferritins are also involved in the protection against oxidative stress during germination and appear to take part in the integrated iron homeostasis establishment. Taken together, my work showed that Arabidopsis ferritins are essential factors that integrate iron and redox homeostasis, while they do not constitute a major iron source for development. (ii) To study ferritin regulation in A. thaliana, the characterization of mutants in the three genes encoding aconitase permitted us to demonstrate that the IRP/IRE system does not occur in the regulation of iron metabolism in plants. Nevertheless, AtFer1 mRNA stability studies have revealed that iron treatment leads to the destabilization of the AtFer1 mRNA. We identified the presence of DST sequences, characterized as mRNA stability determinant, in the 3'-UTR of AtFer1 mRNA. Using chimeric constructs in which the AtFer1 3'-UTR or the AtFer1 3'-UTR with a mutated DST sequence were fused downstream of reporter genes, we have shown that the DST sequence in the 3'-UTR of AtFer1 is functional and sufficient for the iron-dependent mRNA degradation. Using dst1 and dst2 mutants, which are unable to destabilize transcript via DST sequences, we have shown that the DST1 and DST2 gene products, acting in trans in the DST-dependent degradation pathway, are involved in the degradation of AtFer1. Therefore, in addition to the transcriptional regulation described so far, iron is also involved in DST-dependent post-transcriptional regulation of AtFer1 expression. In conclusion, my work has shown that Arabidopsis ferritins are essential elements, which prevent iron toxicity by maintaining a proper labile iron level into the cell, and that sophisticated mechanisms, involving transcriptional and post-transcriptional regulations, permit the tight adjustment of the ferritin accumulation required for the optimal effectiveness of this systemMONTPELLIER-BU Sciences (341722106) / SudocSudocFranceF