In vitro reconstitution of minimal human centrosomes.

ABSTRACT: CDK5RAP2/CEP215 is a key pericentriolar material (PCM) protein that recruits microtubule-nucleating factors at human centrosomes. Using an in vitro reconstitution system, we show that CDK5RAP2 is sufficient to form micron-scale scaffolds around a nanometer-scale nucleator in a PLK-1-regulated manner. CDK5RAP2 assemblies recruited and activated gamma tubulin ring complexes (γ-TuRCs) which, in the presence of α/β tubulin, generated microtubule asters. We found that F75 in CDK5RAP2 is partially needed to recruit γ-TuRC yet is indispensable for γ-TuRC activation. Furthermore, our system recapitulated key features of centrosome-amplified cancer cells. CDK5RAP2 scaffolds selectively recruited the molecular motor KifC1/HSET, which enhanced concentration of α/β tubulin, microtubule polymeri

SUBMITTER: Rios MU 

PROVIDER: S-EPMC11870475 | biostudies-literature | 2025 Feb

REPOSITORIES: biostudies-literature