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Phase separation of hnRNP A1 upon specific RNA-binding observed by magnetic resonance

View ORCID ProfileIrina Ritsch, Elisabeth Lehmann, View ORCID ProfileLeonidas Emmanouilidis, View ORCID ProfileMaxim Yulikov, View ORCID ProfileFrédéric Allain, View ORCID ProfileGunnar Jeschke
doi: https://doi.org/10.1101/2022.03.21.485092
Irina Ritsch
aLaboratory of Physical Chemistry, Department of Chemistry and Applied Bioscience, ETH Zurich, Vladimir-Prelog-Weg 2, 8093 Zurich, Switzerland
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Elisabeth Lehmann
bInstitute of Biochemistry, Department of Biology, ETH Zurich, Hönggerbergring 64, 8093 Zürich, Switzerland
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Leonidas Emmanouilidis
bInstitute of Biochemistry, Department of Biology, ETH Zurich, Hönggerbergring 64, 8093 Zürich, Switzerland
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Maxim Yulikov
aLaboratory of Physical Chemistry, Department of Chemistry and Applied Bioscience, ETH Zurich, Vladimir-Prelog-Weg 2, 8093 Zurich, Switzerland
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Frédéric Allain
bInstitute of Biochemistry, Department of Biology, ETH Zurich, Hönggerbergring 64, 8093 Zürich, Switzerland
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Gunnar Jeschke
aLaboratory of Physical Chemistry, Department of Chemistry and Applied Bioscience, ETH Zurich, Vladimir-Prelog-Weg 2, 8093 Zurich, Switzerland
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  • For correspondence: gjeschke@ethz.ch
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Abstract

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 Statement

The authors have declared no competing interest.

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Posted March 21, 2022.
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Phase separation of hnRNP A1 upon specific RNA-binding observed by magnetic resonance
Irina Ritsch, Elisabeth Lehmann, Leonidas Emmanouilidis, Maxim Yulikov, Frédéric Allain, Gunnar Jeschke
bioRxiv 2022.03.21.485092; doi: https://doi.org/10.1101/2022.03.21.485092
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Phase separation of hnRNP A1 upon specific RNA-binding observed by magnetic resonance
Irina Ritsch, Elisabeth Lehmann, Leonidas Emmanouilidis, Maxim Yulikov, Frédéric Allain, Gunnar Jeschke
bioRxiv 2022.03.21.485092; doi: https://doi.org/10.1101/2022.03.21.485092

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