Single-Molecule Fluorescence Methods to Study Protein-RNA Interactions Underlying Biomolecular Condensates.
Methods Mol Biol
; 2563: 149-160, 2023.
Article
em En
| MEDLINE
| ID: mdl-36227472
ABSTRACT
Many biomolecular condensates, including nucleoli and stress granules, form via dynamic multivalent protein-protein and protein-RNA interactions. These molecular interactions nucleate liquid-liquid phase separation (LLPS) and determine condensate properties, such as size and fluidity. Here, we outline the experimental procedures for single-molecule fluorescence experiments to probe protein-RNA interactions underlying LLPS. The experiments include single-molecule Förster (Fluorescence) resonance energy transfer (smFRET) to monitor protein-induced conformational changes in the RNA, protein-induced fluorescence enhancement (PIFE) to measure protein-RNA encounters, and single-molecule nucleation experiments to quantify the association and buildup of proteins on a nucleating RNA. Together, these experiments provide complementary approaches to elucidate a molecular view of the protein-RNA interactions that drive ribonucleoprotein condensate formation.
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MEDLINE
Assunto principal:
RNA
/
Condensados Biomoleculares
Idioma:
En
Ano de publicação:
2023
Tipo de documento:
Article