Molecular insights into the interaction between a disordered protein and a folded RNA.

Autor:	Mitra R; Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109, USA.; Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA., Usher ET; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, USA.; Center for Biomolecular Condensates (CBC), Washington University in St. Louis, St. Louis, MO, USA., Dedeoğlu S; Centre de Résonance Magnétique Nucléaire à Très Hauts Champs, (CRMN), UMR 5082, CNRS, ENS Lyon, UCBL, Université de Lyon, 69100 Villeurbanne, France., Crotteau MJ; Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109, USA.; Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA., Fraser OA; Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA., Yennawar NH; The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA 16802, USA., Gadkari VV; Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA., Ruotolo BT; Department of Chemistry, University of Michigan, Ann Arbor, MI 48109, USA., Holehouse AS; Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, USA.; Center for Biomolecular Condensates (CBC), Washington University in St. Louis, St. Louis, MO, USA., Salmon L; Centre de Résonance Magnétique Nucléaire à Très Hauts Champs, (CRMN), UMR 5082, CNRS, ENS Lyon, UCBL, Université de Lyon, 69100 Villeurbanne, France., Showalter SA; Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA.; Department of Chemistry, The Pennsylvania State University, University Park, PA 16802, USA., Bardwell JCA; Howard Hughes Medical Institute, University of Michigan, Ann Arbor, MI 48109, USA.; Department of Molecular, Cellular, and Developmental Biology, University of Michigan, Ann Arbor, MI 48109, USA.
Jazyk:	angličtina
Zdroj:	BioRxiv : the preprint server for biology [bioRxiv] 2024 Jun 12. Date of Electronic Publication: 2024 Jun 12.
DOI:	10.1101/2024.06.12.598678
Abstrakt:	Intrinsically disordered protein regions (IDRs) are well-established as contributors to intermolecular interactions and the formation of biomolecular condensates. In particular, RNA-binding proteins (RBPs) often harbor IDRs in addition to folded RNA-binding domains that contribute to RBP function. To understand the dynamic interactions of an IDR-RNA complex, we characterized the RNA-binding features of a small (68 residues), positively charged IDR-containing protein, SERF. At high concentrations, SERF and RNA undergo charge-driven associative phase separation to form a protein- and RNA-rich dense phase. A key advantage of this model system is that this threshold for demixing is sufficiently high that we could use solution-state biophysical methods to interrogate the stoichiometric complexes of SERF with RNA in the one-phase regime. Herein, we describe our comprehensive characterization of SERF alone and in complex with a small fragment of the HIV-1 TAR RNA (TAR) with complementary biophysical methods and molecular simulations. We find that this binding event is not accompanied by the acquisition of structure by either molecule; however, we see evidence for a modest global compaction of the SERF ensemble when bound to RNA. This behavior likely reflects attenuated charge repulsion within SERF via binding to the polyanionic RNA and provides a rationale for the higher-order assembly of SERF in the context of RNA. We envision that the SERF-RNA system will lower the barrier to accessing the details that support IDR-RNA interactions and likewise deepen our understanding of the role of IDR-RNA contacts in complex formation and liquid-liquid phase separation. Competing Interests: DECLARATION OF INTERESTS The authors declare that they have no competing financial interests.
Databáze:	MEDLINE
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