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At the transition from prophase to metaphase, there is a reduction in cohesion along the chromosomal arms, resulting in the resolution of sister chromatids. However, residual cohesin connections remain to hold the sister chromatids together until the transition from metaphase to anaphase. The residual connection prevents any premature separation of sister chromatids, blocking the risks of aneuploidy within the daughter cells.
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"Off-pore" nucleoporins relocalize heterochromatic breaks through phase separation.

Chiara Merigliano1, Taehyun Ryu2, Colby D See1

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Summary

Nuclear phase separation by Nup98 protein mobilizes DNA repair sites to the nuclear periphery. This prevents harmful recombination and ensures safe homologous recombination (HR) repair, maintaining genome integrity.

Keywords:
Heterochromatin repairNup88Nup98Sec13condensatesdouble-strand break repairhomologous recombinationnuclear architecturephase separationrepair dynamics

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Area of Science:

  • Cell biology
  • Molecular genetics
  • Genomics

Background:

  • Membraneless compartments form via phase separation in the nucleus, organizing heterochromatin and DNA repair foci.
  • Pericentromeric heterochromatin contains repetitive sequences susceptible to aberrant recombination.
  • Safe homologous recombination (HR) repair of these sequences necessitates movement to the nuclear periphery prior to Rad51 recruitment.

Purpose of the Study:

  • To investigate the mechanisms initiating the mobilization of heterochromatic repair sites.
  • To elucidate the role of phase separation in these nuclear dynamics.
  • To understand the contribution of nucleoporins to heterochromatin repair and genome stability.

Main Methods:

  • Recruitment assays for Nup98 at heterochromatic repair sites.
  • Analysis of nucleoporin relocalization dynamics (Sec13, Nup88).
  • Investigating the role of Smc5/6 complex and SUMOylation.
  • Assessing the impact of Nup98 phase separation properties on repair site mobilization and Rad51 exclusion.
  • Evaluating heterochromatin repair and chromosome integrity upon pathway disruption.

Main Results:

  • Nup98 nucleoporin is recruited to heterochromatic repair sites before relocalization via Sec13 or Nup88.
  • Nup98 recruitment occurs downstream of the Smc5/6 complex and SUMOylation.
  • Nup98's phase separation properties are essential and sufficient for mobilizing repair sites and excluding Rad51.
  • Disruption of this pathway leads to heterochromatin repair defects and widespread chromosome rearrangements.

Conclusions:

  • Nup98-mediated phase separation drives the mobilization of DNA repair sites to the nuclear periphery.
  • This process is crucial for preventing aberrant recombination and ensuring faithful HR repair in heterochromatin.
  • Nucleoporins play a novel "off-pore" role in nuclear dynamics and maintaining genome integrity through phase separation.