Obiettivo
The role of metabolic enzymes in the post-transcriptional control of gene expression is barely explored, with only a few cases deeply studied so far, such as aconitase 1 in iron metabolism or GAIT complex control of pro-inflammatory genes. Here, we propose to identify metabolic enzymes that interact specifically with mRNA by unbiased proteomic and transcriptomic approaches and to investigate their potential roles as gene expression regulators. Metabolic enzymes able to bind mRNAs will be identified by an effective crosslinking method, followed by oligo d(T) isolation and mass spectrometry in both human and yeast systems. Following this first screening, we will select three conserved key metabolic enzymes involved in the regulation of different metabolic pathways for further in depth exploration. Using cross-linking and immunoprecipitation assays followed by deep sequencing, we will identify the specific pool of mRNAs bound to each selected metabolic enzyme. Next, we will determine the RNA motif and the protein domain responsible for the interaction with biochemical and bioinformatic tools. Finally, the effect of overexpression and depletion of these metabolic enzymes in the post-transcriptional control of their target mRNAs will be analyzed. Overall, the proposed project will explore a potential new layer of cell regulation.
Campo scientifico
Argomento(i)
Invito a presentare proposte
FP7-PEOPLE-2009-IEF
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Meccanismo di finanziamento
MC-IEF - Intra-European Fellowships (IEF)Coordinatore
69117 Heidelberg
Germania