Fig. 2: DONSON maintains centriole engagement through the translocation of Cdc6 from the nucleus to the centrosome.

a Subcellular localization of overexpressed DONSON and Cdc6 in G1 and S/G2 phases. HeLa-Tet3G cells were treated with 1 μg/mL Doxycycline (Dox) for 24 h. b HeLa cells were fixed 24 h after synchronization in G1 phase using 10 µM PHA767491 (Cdc7i) or 4 h after release. c Cdc6 localization at centrioles in S phase in control or DONSON-depleted cells. d Quantification of the normalized signal intensity of Cdc6 at each centriole observed in (c). n  =  50 from three independent experiments. e Inducible expression of Cdc6 upon DONOSN depletion. HeLa-Tet3G-mNG-Cdc6 cells were treated with siRNA for 48 h, followed by 1 μg/mL Dox treatment for 24 h. f Histograms represent the frequency of interphase cells with > 2 C-Nap1 foci observed in (e). n  =  3 independent experiments, 30 cells for each. g List of proteins previously reported to be involved in centriole engagement or disengagement. h Precocious centriole disengagement was inhibited upon co-depletion of Cdh1 and DONSON/Cdc6. i, j Histograms represent the frequency of interphase cells with >2 C-Nap1 foci observed in (h). n  =  3 independent experiments, 50 cells for each. k Cdh1 localization at centrosomes in cells treated with siControl or siCdc6. l Inducible expression of Cdc6 upon DONOSN depletion. HeLa-Tet3G-mRuby3-Cdc6 cells were treated with siRNA for 48 h, followed by 1 μg/mL Dox treatment for 24 h. m Quantification of the normalized signal intensity of Cdh1 at each centriole with the indicated treatment. n = 50 from three independent experiments. n Schematic illustration of the phenotype of DONSON depletion. The depletion of DONSON leads to the mislocalization of Cdc6 at the centrosomes during the S phase, where Cdc6 inhibits the premature recruitment of Cdh1. All scale bars, 5 μm. Values are mean percentages ± s.d. Tukey’s multiple comparison test was used in (i), Two-tailed, unpaired Student’s t-test was used in (f) and (j), Mann–Whitney U test was used in (d), and Kruskal–Wallis test was used in (m) to obtain P-value. Source data are provided as a Source Data file.