Barrier properties of Nup98 FG phases ruled by FG motif identity and inter-FG spacer length

Sheung Chun Ng, Abin Biswas, Trevor Huyton, Jürgen Schünemann, Simone Reber and Dirk Görlich ()
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Sheung Chun Ng: Max Planck Institute for Multidisciplinary Sciences
Abin Biswas: Humboldt-Universität zu Berlin
Trevor Huyton: Max Planck Institute for Multidisciplinary Sciences
Jürgen Schünemann: Max Planck Institute for Multidisciplinary Sciences
Simone Reber: Humboldt-Universität zu Berlin
Dirk Görlich: Max Planck Institute for Multidisciplinary Sciences

Nature Communications, 2023, vol. 14, issue 1, 1-17

Abstract: Abstract Nup98 FG repeat domains comprise hydrophobic FG motifs linked through uncharged spacers. FG motifs capture nuclear transport receptors (NTRs) during nuclear pore complex (NPC) passage, confer inter-repeat cohesion, and condense the domains into a selective phase with NPC-typical barrier properties. We show that shortening inter-FG spacers enhances cohesion, increases phase density, and tightens such barrier - all consistent with a sieve-like phase. Phase separation tolerates mutating the Nup98-typical GLFG motifs, provided domain-hydrophobicity remains preserved. NTR-entry, however, is sensitive to (certain) deviations from canonical FG motifs, suggesting co-evolutionary adaptation. Unexpectedly, we observed that arginines promote FG-phase-entry apparently also by hydrophobic interactions/ hydrogen-bonding and not just through cation-π interactions. Although incompatible with NTR·cargo complexes, a YG phase displays remarkable transport selectivity, particularly for engineered GFPNTR-variants. GLFG to FSFG mutations make the FG phase hypercohesive, precluding NTR-entry. Extending spacers relaxes this hypercohesion. Thus, antagonism between cohesion and NTR·FG interactions is key to transport selectivity.

Date: 2023
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DOI: 10.1038/s41467-023-36331-4

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