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Box C/D snoRNPs: solid-state NMR fingerprint of an early-stage 50 kDa assembly intermediate

Abstract : Many cellular functions rely on stable protein-only or protein-RNA complexes. Deciphering their assembly mechanism is a key question in cell biology. We here focus on box C/D small nucleolar ribonucleoproteins involved in ribosome biogenesis. The mature particles contain four core proteins and a guide RNA. Despite their relatively simple composition, these particles don't self-assemble in eukaryote and the production of a native and functional particle requires a large number of transient other proteins, called assembly factors. We present here 13C and 15N solid-state NMR assignment of yeast 126-residue core protein Snu13 in the context of its 50 kDa pre-complex with assembly factors Rsa1p:Hit1p. In this sample, only one third of the protein is labelled, leading to a low sensitivity. We could nevertheless obtain assignment data for 91% of the residues. Secondary structure derived from our assignments shows that Snu13p overall structure is maintained in the context of the complex. Chemical shift perturbations are analysed to evaluate Snu13p conformational changes and interaction interface upon binding to its partner proteins. While indirect perturbations are observed in the hydrophobic core, we find other good candidate residues belonging to the interaction interface. We describe the role of some Snu13p N-terminal and C-terminal residues, not identified in previous structural studies. These preliminary results will serve as a basis for future interaction studies, especially by adding RNA, to decipher box C/D snoRNP particles assembly pathway.
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Contributor : JULIE GONNET Connect in order to contact the contributor
Submitted on : Thursday, December 10, 2020 - 10:36:14 AM
Last modification on : Saturday, September 24, 2022 - 3:10:05 PM
Long-term archiving on: : Thursday, March 11, 2021 - 7:00:04 PM


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Marie-Eve Chagot, Marc Quinternet, Clémence Jacquemin, Xavier Manival, Carole Gardiennet. Box C/D snoRNPs: solid-state NMR fingerprint of an early-stage 50 kDa assembly intermediate. Biomolecular NMR Assignments, Springer, 2020, 14 (1), pp.131-140. ⟨10.1007/s12104-020-09933-y⟩. ⟨hal-02480749⟩



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