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  • Differential Requirements for Centrioles in Mitotic Centrosome Growth and Maintenance.

Differential Requirements for Centrioles in Mitotic Centrosome Growth and Maintenance.

Developmental cell (2019-07-16)
Gabriela Cabral, Triin Laos, Julien Dumont, Alexander Dammermann
ABSTRACT

Centrosomes, the predominant sites of microtubule nucleation and anchorage, coordinate spindle assembly and cell division in animal cells. At the onset of mitosis, centrioles accumulate microtubule-organizing pericentriolar material (PCM) in a process termed centrosome maturation. To what extent centrosome maturation depends on the continued activity of mitotic regulators or the presence of centrioles has hitherto been unclear. Using the C. elegans early embryo, we show that PCM expansion requires the Polo-like kinase PLK-1 and CEP192 (SPD-2 in C. elegans), but not its upstream regulator Aurora A (AIR-1), while maintenance of the PCM polymer depends exclusively on PLK-1. SPD-2 and PLK-1 are highly concentrated at centrioles. Unexpectedly, laser microsurgery reveals that while centrioles are required for PCM recruitment and centrosome structural integrity they are dispensable for PCM maintenance. We propose a model whereby centrioles promote centrosome maturation by recruiting PLK-1, but subsequent maintenance occurs via PLK-1 acting directly within the PCM.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Nocodazole, Inhibitor of mitosis.
Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, clone DM1A, ascites fluid
Sigma-Aldrich
clasto-Lactacystin β-Lactone, A highly specific, cell-permeable, and irreversible 20S proteasome inhibitor.
Roche
cOmplete, Mini, EDTA-free Protease Inhibitor Cocktail, Protease Inhibitor Cocktail Tablets provided in a glass vial, Tablets provided in a glass vial