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Updated: Apr 21, 2026

In Vivo Proximity Biotinylation for Protein Interaction Studies in Paramecium tetraurelia
Published on: September 12, 2025
A developmental condensin I complex assists the Paramecium PiggyMac domesticated transposase during programmed DNA
Thomas Balan1, Mélanie Bazin-Gélis2, Marc Guérineau2
1Université Paris Cité, CNRS, Institut Jacques Monod, Paris 75013, France.
Abstract:
Prokaryotes and eukaryotes use diverse strategies to cope with invading mobile genetic elements, including programmed DNA elimination (PDE). In the ciliate Paramecium, elimination of transposable elements and their relics requires the PiggyMac (Pgm) endonuclease and its five PgmL partners, yet how this machinery is targeted to cleavage sites remains unclear. Here, we identified condensin I subunits in the proximity proteomes of Pgm and PgmL4. We show that they belong to a condensin complex that is essential for PDE and localizes to developing somatic nuclei. Depleting the development-specific subunits of this complex blocks DNA elimination, phenocopying a Pgm depletion. Developmental condensin is required for the correct nuclear localization of Pgm and some of the PgmLs. Moreover, Pgm and these PgmLs coimmunoprecipitate with condensin I. Our findings uncover functional and physical interactions between a eukaryotic DNA cleavage machinery and a specialized condensin complex that is critical for PDE in a nondividing nucleus.
Insights
A specialized condensin complex is essential for programmed DNA elimination (PDE) in Paramecium. This developmental condensin targets the PiggyMac (Pgm) endonuclease machinery to DNA cleavage sites for removing mobile genetic elements.
Area of Science:
- Molecular Biology
- Genetics
- Cell Biology
Background:
- Eukaryotes employ programmed DNA elimination (PDE) to manage mobile genetic elements.
- The PiggyMac (Pgm) endonuclease and its PgmL partners are crucial for PDE in Paramecium, but their targeting mechanism is unknown.
Purpose of the Study:
- To elucidate the targeting mechanism of the Pgm endonuclease machinery during programmed DNA elimination (PDE).
- To investigate the role of condensin I in the DNA elimination process.
Main Methods:
- Proximity proteome analysis of Pgm and PgmL4.
- Depletion of specific condensin I subunits.
- Immunoprecipitation assays.
- Subnuclear localization studies.
Main Results:
- Condensin I subunits were identified in proximity to Pgm and PgmL4, forming a complex essential for PDE.
- This developmental condensin localizes to developing somatic nuclei and is required for Pgm and PgmL localization.
- Depletion of developmental condensin subunits phenocopies Pgm depletion, blocking DNA elimination.
- Functional and physical interactions between condensin I and the Pgm/PgmL DNA cleavage machinery were demonstrated.
Conclusions:
- A specialized condensin I complex plays a critical role in targeting the Pgm endonuclease machinery for programmed DNA elimination (PDE).
- This interaction is vital for managing transposable elements in the somatic nucleus of Paramecium.
- Findings reveal a novel link between nuclear architecture and DNA cleavage machinery in genome defense.
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