Reconstituting centriole biogenesis on an engineered cellular platform uncovers organelle assembly principles
Centriole copy number is tightly regulated, with one procentriole emanating from a torus surrounding each pre-existing centriole. Which proteins are sufficient to generate a full-fledged organelle is unclear. We address this question by engineering a high valency low copy number phase-separated droplet platform to concentrate proteins in an ectopic cellular location. We establish that droplet…
Scientists have engineered a cellular platform to uncover the principles of centriole assembly. Centrioles have a tightly regulated copy number, with one procentriole emerging from a torus surrounding each pre-existing centriole. The identity of the proteins responsible for generating a complete organelle is unknown. To investigate this, researchers developed a high valency low copy number phase-separated droplet platform to concentrate proteins in an artificial location.
They found that targeting the torus protein Cep63 to the droplet is sufficient to initiate procentriole assembly. When the interaction involving STIL is relieved or pre-existing centrioles are absent, ectopic procentrioles mature. These synthetic centrioles detach from the droplet during mitosis, organize additional spindle poles, and trigger procentriole formation in the next cell cycle, confirming the reconstitution of synthetic centrioles.
Additionally, the number of procentrioles scales with the surface area of the droplet platform, indicating that this constitutes a closed circuit mechanism governing organelle number homeostasis.
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