Regulated assembly of a supramolecular centrosome scaffold in vitro

• Published in: Science (American Association for the Advancement of Science) Vol. 348; no. 6236; pp. 808 - 812
• Main Authors: Woodruff, Jeffrey B., Wueseke, Oliver, Viscardi, Valeria, Mahamid, Julia, Ochoa, Stacy D., Bunkenborg, Jakob, Widlund, Per O., Pozniakovsky, Andrei, Zanin, Esther, Bahmanyar, Shirin, Zinke, Andrea, Hong, Sun Hae, Decker, Marcus, Baumeister, Wolfgang, Andersen, Jens S., Oegema, Karen, Hyman, Anthony A.
• Format: Journal Article
• Language: English
• Published: Washington American Association for the Advancement of Science 15-05-2015; The American Association for the Advancement of Science
• Subjects: Annan medicinsk grundvetenskap; Assembly; Caenorhabditis elegans; Cellular biology; Coiling; Cytology; In vitro testing; Molecular biology; Networks; Other Basic Medicine; Polymerization; Proteins; Regulators; Scaffolds
• ISSN: 0036-8075; 1095-9203
• DOI: 10.1126/science.aaa3923

The centrosome organizes microtubule arrays within animal cells and comprises two centrioles surrounded by an amorphous protein mass called the pericentriolar material (PCM). Despite the importance of centrosomes as microtubule-organizing centers, the mechanism and regulation of PCM assembly are not well understood. In Caenorhabditis elegans, PCM assembly requires the coiled-coil protein SPD-5. We found that recombinant SPD-5 could polymerize to form micrometer-sized porous networks in vitro. Network assembly was accelerated by two conserved regulators that control PCM assembly in vivo, Polo-like kinase-1 and SPD-2/Cep192. Only the assembled SPD-5 networks, and not unassembled SPD-5 protein, functioned as a scaffold for other PCM proteins. Thus, PCM size and binding capacity emerge from the regulated polymerization of one coiled-coil protein to form a porous network.