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  • Drosophila Tubulin-Specific Chaperone E Recruits Tubulin around Chromatin to Promote Mitotic Spindle Assembly.

Drosophila Tubulin-Specific Chaperone E Recruits Tubulin around Chromatin to Promote Mitotic Spindle Assembly.

Current biology : CB (2020-12-02)
Mathieu Métivier, Emmanuel Gallaud, Alexandre Thomas, Aude Pascal, Jean-Philippe Gagné, Guy G Poirier, Denis Chrétien, Romain Gibeaux, Laurent Richard-Parpaillon, Christelle Benaud, Régis Giet
ABSTRACT

Proper assembly of mitotic spindles requires microtubule nucleation not only at the centrosomes but also around chromatin. In this study, we found that the Drosophila tubulin-specific chaperone dTBCE is required for the enrichment of tubulin in the nuclear space after nuclear envelope breakdown and for subsequent promotion of spindle microtubule nucleation. These events depend on the CAP-Gly motif found in dTBCE and are regulated by Ran and lamin proteins. Our data suggest that during early mitosis, dTBCE and nuclear pore proteins become enriched in the nucleus, where they interact with the Ran GTPase to promote dynamic tubulin enrichment. We propose that this novel mechanism enhances microtubule nucleation around chromatin, thereby facilitating mitotic spindle assembly.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Anti-α-Tubulin antibody, Mouse monoclonal, clone DM1A, purified from hybridoma cell culture
Roche
Anti-GFP, from mouse IgG1κ (clones 7.1 and 13.1)
Sigma-Aldrich
Mineral oil, light oil, (neat)
Sigma-Aldrich
Anti-phospho-Histone H3 (Ser10) Antibody, Mitosis Marker, Upstate®, from rabbit
Sigma-Aldrich
Anti-Tubulin Antibody, clone YL1/2, clone YL1/2, Chemicon®, from rat