Pérez-Ràfols, A.* ; Pérez-Ropero, G.* ; Cerofolini, L.* ; Sperotto, L. ; Roca-Martínez, J.* ; Higuera-Rodriguez, R.A.* ; Russomanno, P.* ; Kaiser, W.J.* ; Vranken, W.F.* ; Danielson, U.H.* ; Provenzani, A.* ; Martelli, T.* ; Sattler, M. ; Buijs, J.* ; Fragai, M.*
Deciphering the RNA recognition by Musashi-1 to design protein and RNA variants for in vitro and in vivo applications.
Nucleic Acids Res. 53:gkaf741 (2025)
The Human Musashi-1 (MSI-1) is an RNA-binding protein that recognizes (G/A)U1-3AGU and UAG sequences in diverse RNAs through two RNA Recognition Motif (RRM) domains and regulates the fate of target RNA. Here, we have combined structural biology and computational approaches to analyse the binding of the RRM domains of human MSI-1 with single-stranded and structured RNA ligands. We have used our recently developed computational tool RRMScorer to design a set of substitutions in the MSI-1 protein and the investigated RNA strands to modulate the binding affinity and selectivity. The in silico predictions of the designed protein-RNA interactions are assessed by nuclear magnetic resonance and surface plasmon resonance. These experiments have also been used to study the competition of the two RRM domains of MSI-1 for the same binding site within linear and harpin RNA. Our experimental results shed light on MSI-RNA interactions, thus opening the way for the development of new biomolecules for in vitro and in vivo studies and downstream applications.
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Publication type
Article: Journal article
Document type
Scientific Article
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Keywords
Messenger-rna; Binding Proteins; Gene-expression; Motif; Relaxation; Target; Nmr
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Language
english
Publication Year
2025
Prepublished in Year
0
HGF-reported in Year
2025
ISSN (print) / ISBN
0305-1048
e-ISSN
1362-4962
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Volume: 53,
Issue: 15,
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Article Number: gkaf741
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Oxford University Press
Publishing Place
Great Clarendon St, Oxford Ox2 6dp, England
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Reviewing status
Peer reviewed
POF-Topic(s)
30203 - Molecular Targets and Therapies
Research field(s)
Enabling and Novel Technologies
PSP Element(s)
G-503000-001
Grants
European Union NextGenerationEU
Next Generation EU
Marie Skłodowska-Curie Innovative Training Network (MSCA-ITN) RNAct
Copyright
Erfassungsdatum
2025-10-10