Emanuela Marchese
| Deciphering the impact of phosphorylation on HuR functional domains through molecular dynamics simulations |
| Marchese Emanuela1, Paravati Maria Rosaria1, Ambrosio Francesca Alessandra1, Castoria Gabriella2, Tesei Anna3, Costa Giosuè1, Collina Simona4, and Alcaro Stefano1,5 1 Dipartimento di Scienze della Salute, Università degli Studi “Magna Græcia” di Catanzaro, Campus “S. Venuta”, Viale Europa, 88100, Catanzaro, Italy 2 Dipartimento di Medicina di Precisione, Università della Campania “L. Vanvitelli”, Naples, Italy 3 Biosciences Laboratory, IRCCS Istituto Scientifico Romagnolo per lo Studio e la Cura dei Tumori (IRST) “Dino Amadori”, I-47014 Meldola, Italy 4 Department of Drug Sciences, University of Pavia, Via Taramelli 12, 27100, Pavia, Italy 5 Associazione CRISEA – Centro di Ricerca e Servizi Avanzati per l’Innovazione Rurale, località Condoleo di Belcastro Catanzaro, Italy |
| Abstract RNA-binding proteins (RBPs) are key regulators of post-transcriptional gene expression and promising therapeutic targets in several pathologies [1]. Human antigen R (HuR, encoded by ELAVL1) is a well-characterized RBP that modulates the stability and translation of mRNAs involved in oncogenesis and inflammation. HuR activity is tightly controlled by post-translational modifications, particularly phosphorylation, which affects RNA-binding affinity and subcellular localization. Specifically, phosphorylation sites located within the RRM1 and RRM2 domains (S88, S100, T118) have been shown to directly influence HuR interaction with target transcripts, promoting dissociation from specific mRNAs and thereby modulating the stability of transcripts involved in proliferation and survival pathways. In contrast, phosphorylation events in the RRM3 domain and hinge region (S221, S318) regulate HuR nucleocytoplasmic shuttling [2]. Despite these insights, the molecular mechanisms by which site-specific phosphorylation alters HuR’s conformational dynamics and RNA recognition remain poorly understood. Here, we first employed GRID-based interaction energy studies to map favorable phosphorylation sites within HuR functional domains [3]. This mapping guided subsequent comparative MD simulations, performed to investigate the structural effects of phosphorylation on HuR variants [4]. By characterizing the protein–RNA interface and global flexibility, we aim to provide a mechanistic basis for how these modifications modulate HuR function. This work could establish a structural framework for the rational design of selective HuR modulators, offering new avenues for therapeutic intervention in oncology. |
| References [1] Wutikeli, H.; Xie, T.; Xiong, W.; Shen, Y. ELAV/Hu RNA-binding protein family: key regulators in neurological disorders, cancer, and other diseases. RNA biology 2025, 22(1), 1–11. https://doi.org/10.1080/15476286.2025.2471133 [2] Grammatikakis, I.; Abdelmohsen, K.; Gorospe, M. Posttranslational Control of HuR Function. WIREs RNA 2016, 8(1). https://doi.org/10.1002/wrna.1372 [3] Goodford, P. J. A computational procedure for determining energetic targets for specificity in computer-aided drug design. Journal of Medicinal Chemistry 1985, 28(7), 849-857. https://doi.org/10.1021/jm00145a002 [4] Schrödinger Release 2024-2: Maestro; Schrödinger LLC: New York, NY, USA, 2024 |