Structural studies on mRNA recognition by FMRP and TDP-43, two RNA-binding proteins associated with neuronal diseases

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The deregulation of posttranscriptional regulatory networks by genetic mutation, deletion or overexpression of RNA-binding proteins results in disease, yet their target RNAs and their modes of regulation are largely unknown. Many neurodegenerative conditions are caused by altered RBP expression that perturbs post-transcriptional regulatory networks [1]. Examples include Fragile-X-syndrome (FXS) [2] and amyotrophic lateral sclerosis (ALS) [3]. This research project aims to understand the recognition of mRNA targets to deduce the regulatory principles of two disease-associated RBPs, one of which (FMRP) is associated with the FXS and particular types of autism, whereas mutations in the other (TDP-43) are associated with ALS. Molecular RNA targets of these RBPs have been identified in the host's group by using PAR-CLIP (photoactivatable ribonucleotide crosslinking and immunoprecipitation) in combination with high-throughput sequencing. This method allows an efficient identification of cellular RNA targets and the determination of RNA recognition elements from their consensus sequences. The RBPs will be analyzed in the presence and absence of their biologically relevant RNA targets as well as further interaction partners using biophysical and structural-biology methods. This approach will go beyond analyses of short RNA fragments with isolated RNA-binding domains. Results on the structure and folding states of the RBPs in the presence and absence of their ligands will allow the generation of hypotheses of their molecular function and regulation, which will be investigated using in vitro models and living cells. A further understanding of RBPs at the molecular level as well as in a cellular environment may contribute to the development of specific pharmacological interventions for RBP-involving diseases.