RT Journal Article
SR Electronic
T1 Phase separation of hnRNP A1 upon specific RNA-binding observed by magnetic resonance
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2022.03.21.485092
DO 10.1101/2022.03.21.485092
A1 Ritsch, Irina
A1 Lehmann, Elisabeth
A1 Emmanouilidis, Leonidas
A1 Yulikov, Maxim
A1 Allain, Frédéric
A1 Jeschke, Gunnar
YR 2022
UL http://biorxiv.org/content/early/2022/03/21/2022.03.21.485092.abstract
AB Interaction of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) with specific single-stranded RNA and its relation to liquid-liquid phase separation were investigated in vitro by magnetic resonance based on site-directed spin labelling. An ensemble model of free hnRNP A1 in the absence of RNA was derived from distance distributions between spin labelled sites and small angle X-ray scattering. This model revealed a compact state of the low-complexity domain and interaction of this domain with the RNA recognition motifs. Paramagnetic relaxation enhancement NMR spectroscopy confirmed this interaction. The addition of RNA to dispersed solutions of hnRNP A1 induced phase separation, observed by formation of liquid droplets. The phase separation depended on the RNA concentration and sequence, with continuous wave EPR spectroscopy showing that local protein dynamics is affected by point mutations in the RNA sequence. We propose that an interplay of sequence-specific RNA binding and phase transition serves as a regulatory mechanism for RNA segregation in the stress response of cells.Competing Interest StatementThe authors have declared no competing interest.