PLK4 promotes centriole duplication by phosphorylating STIL to link the procentriole cartwheel to the microtubule wall

Abstract

Centrioles play critical roles in organizing the assembly of the mitotic spindle and templating the formation of primary cilia. Centriole duplication occurs once per cell cycle and is regulated by Polo-like kinase 4 (PLK4). Although significant progress has been made in understanding centriole composition, we have limited knowledge of how PLK4 activity controls specific steps in centriole formation. Here, we show that PLK4 phosphorylates its centriole substrate STIL on a conserved site, S428, to promote STIL binding to CPAP. This phospho-dependent binding interaction is conserved in Drosophila and facilitates the stable incorporation of both STIL and CPAP into the centriole. We propose that procentriole assembly requires PLK4 to phosphorylate STIL in two different regions: phosphorylation of residues in the STAN motif allow STIL to bind SAS6 and initiate cartwheel assembly, while phosphorylation of S428 promotes the binding of STIL to CPAP, linking the cartwheel to microtubules of the centriole wall.

The Singularity of the Drosophila Male Germ Cell Centriole: The Asymmetric Distribution of Sas4 and Sas6

“…Otherwise, the asymmetric accumulation of Sas6 might reflect the specific interactions with different centriolar proteins. Remarkably, the conserved centriole duplication factor Ana2 that interacts with Sas-6 and plays a main role in its recruitment [56][57][58][59][60][61] is also more concentrated to the daughter centrioles in primary spermatocytes [39]. However, whereas Sas-6 is restricted to the proximal region of the centriole, Ana2 is localized along the whole centriole length [39].…”

TRIM37 prevents formation of centriolar protein assemblies by regulating Centrobin stability

“…(12780-1-AP; Proteintech), 1:1000 mouse anti-Centrobin (ab70448; Abcam), 1:1000 rabbit anti-Centrobin (HPA023321; Atlas Antibodies), 1:1000 rabbit anti-CEP135 (ab75005; Abcam), 1:500 rabbit anti-CPAP (Kohlmaier et al, 2009), 1:500 rabbit anti-SPICE (HPA064843, Sigma-Aldrich), 1:8,000 rabbit anti-Ninein (L77), 1:1000 rabbit anti-hPOC5 (Azimzadeh et al, 2009) and 1:1000 rabbit anti-PLK4(KD) (Sillibourne et al, 2010) (both generous gifts from Michel Bornens, Institut Curie, Paris, France), 1:1000 rabbit anti-P-PLK4 (Moyer and Holland, 2019) (generous gifts from Andrew Holland), 1:400 mouse anti-C-Nap (611374; BD Biosciences) 1:2000 rabbit anti-STIL (ab222838; Abcam), 1:1000 rabbit anti-PCNT (ab4448; Abcam), 1:1000 mouse anti-AKAP450 (611518; BD Biosciences), 1:1000 rabbit anti-CDK5Rap2 (06-1398; Millipore), 1:1000 rabbit anti-CEP192…”

Differential turnover of Nup188 controls its levels at centrosomes and role in centriole duplication

“…To double the centrosome number, each of the centrioles must first be duplicated in S-phase by a nascent “pro” centriole that assembles perpendicularly from the parent centriole outer wall (Nigg and Holland, 2018). This is achieved by a highly coordinated series of yet to be fully defined molecular events triggered by the recruitment of polo-like–kinase 4 (Plk4; Bettencourt-Dias et al, 2005; Habedanck et al, 2005; O’Connell et al, 2001) by Cep152 and Cep192 (Cizmecioglu et al, 2010; Hatch et al, 2010; Kim et al, 2013; Park et al, 2014; Sonnen et al, 2013) and its phosphorylation of a network of proteins including STIL, CPAP, and Sas6 (Dammermann et al, 2004; Delattre et al, 2004; Dzhindzhev et al, 2017; Kemp et al, 2004; Kirkham et al, 2003; Kratz et al, 2015; Leidel et al, 2005; Leidel and Gönczy, 2003; Moyer et al, 2015; Moyer and Holland, 2019; Ohta et al, 2014; Pelletier et al, 2004; Stevens et al, 2010; Tang et al, 2011). Sas6 helps form a supramolecular cartwheel-shaped structure that serves as a scaffold for microtubule nucleation (Gönczy, 2012).…”