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Related Experiment Video

Updated: Jul 1, 2025

Reconstitution of Basic Mitotic Spindles in Spherical Emulsion Droplets

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Published on: August 13, 2016

Multivalent coiled-coil interactions enable full-scale centrosome assembly and strength.

Manolo U Rios¹, Małgorzata A Bagnucka¹, Bryan D Ryder²

¹Department of Cell Biology, Department of Biophysics, The University of Texas Southwestern Medical Center, Dallas, TX, USA.

The Journal of Cell Biology

|March 8, 2024

Summary

This summary is machine-generated.

Phosphorylation by PLK-1 opens the SPD-5 scaffold protein, enabling pericentriolar material (PCM) assembly. This process involves multivalent coiled-coil interactions, crucial for mitotic spindle organization and PCM strength.

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Area of Science:

Cell Biology
Molecular Biology
Structural Biology

Background:

The pericentriolar material (PCM) is essential for organizing microtubules and forming the mitotic spindle.
Understanding the molecular mechanisms governing PCM assembly and its mechanical properties is critical.

Purpose of the Study:

To investigate the molecular interactions driving pericentriolar material (PCM) assembly.
To elucidate the role of SPD-5 scaffold protein phosphorylation in PCM formation and mechanics.

Main Methods:

Crosslinking mass spectrometry (XL-MS) to analyze protein multimerization.
Structural analysis of specific protein regions and mutations.
Investigating the impact of microtubule-mediated forces on PCM assembly.

Main Results:

Phosphorylation of SPD-5 by PLK-1 eliminates intramolecular crosslinks, inducing a conformational opening.
SPD-5 multimerization is mediated by interactions between dispersed coiled-coil domains.
Mutations in SPD-5 interacting regions cause PCM assembly defects, partially rescued by reducing microtubule forces, indicating interdependence of assembly and strength.

Conclusions:

PCM assembly and mechanical strength are interdependent processes.
Multivalent coiled-coil interactions between SPD-5 proteins drive PCM size and strength.
Phosphorylation-induced structural changes in SPD-5 are key to initiating PCM assembly.