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C-P.45: Mapping Evolutionary Switches Driving Functional Diversification in AsnC-like Transcription Factors

Authors

Pedro Beltrao
Institute of Molecular Systems Biology, ETH Zürich
Julian Trouillon
Institute of Molecular Systems Biology, ETH Zürich

Keywords

ancestral sequence reconstruction, specificity-determining residues, transcription factor evolution, protein-DNA recognition, effector binding, Lrp/AsnC family
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Deciphering the transcriptional regulatory code requires understanding how a transcription factor's amino acid sequence dictates its DNA-binding specificity. AsnC-like transcription factors (InterPro: IPR019888) form an ancient regulatory superfamily that offers a dense evolutionary sampling for this purpose. Here, we leverage the phylogeny of the AsnC-like family to systematically map this functional diversification. To identify the key changes causing functional diversification, we used the burst after duplication divergence metric with ancestral sequence reconstructions. This approach highlights "evolutionary switches" - residues conserved within subfamilies but different between them, suggesting strong functional importance. By mapping the high-scoring residues onto structural models, we can deduce their specific biological roles: switches at the protein-DNA interface suggest residues that determine specificity, while those at effector-binding or dimerization sites show the evolution of metabolic sensing and complex assembly. Beyond this, we are broadening our analysis to explore the larger factors and timing of these functional shifts. By characterizing the physicochemical nature of the mutations at switch sites, we aim to gain understanding of how specific substitutions alter function. By linking the identified evolutionary switches with known DNA-binding specificities, effector molecules, and the host species' environments, we can deduce the selective pressures that drive regulatory adaptation. Mapping these switches across the phylogeny also allows us to estimate when these adaptations emerged. This evolutionary approach ultimately aims at clarifying how regulatory domains take on novel functions in the AsnC-like transcription factor family, offering a sound basis for understanding adaptation through protein evolution. Co-authors: Pedro Beltrao, Julian Trouillon

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