Fig. 7: Patient-derived CEP112 mutations disrupt phase separation dynamics and impair translational regulation. | Nature Communications

Fig. 7: Patient-derived CEP112 mutations disrupt phase separation dynamics and impair translational regulation.

From: CEP112 coordinates translational regulation of essential fertility genes during spermiogenesis through phase separation in humans and mice

Fig. 7

a RT-PCR analysis of wild-type (WT) and mutant CEP112 in HEK293T cells shows significantly reduced expression of the R76X mutant, while S137L, G90R, and H766R mutants have comparable expression levels to WT. Three independent experiments were performed. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. Source data are provided as a Source Data file. b Western blot analysis of CEP112 proteins in HEK293T cells. Representative blot and quantification of CEP112 relative to GAPDH. Three independent experiments were performed. (Two-sided Student’s t test; error bars, mean ± SEM). Source data are provided as a Source Data file. c–e Structural impact of missense mutations (S137L, G90R, H766R) predicted by computational modeling, with mutated residues highlighted in purple. f Fluorescence microscopy shows WT and mutant CEP112 (S137L, G90R, H766R) forming spherical condensates in HEK293T cells (Scale bar: 10 μm). g Quantification of condensates per cell with WT or mutant CEP112 in HEK293T cells. n = 50 cells per group. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. Source data are provided as a Source Data file. h FRAP analysis of condensates in HEK293T cells reveals that the S137L mutant recovers similarly to WT, while G90R and H766R mutants show slower recovery, indicating altered liquid-like properties. Three independent experiments were performed. Data are presented as mean values ± SEM. Source data are provided as a Source Data file. i TRICK reporter assay in COS-7 cells shows WT CEP112 sequesters CFAP61 mRNA into granules, promoting translation (RFP signal only). Mutant CEP112 disrupts this, with GFP and RFP signals indicating impaired translational regulation (left panel, Scale bar: 10 μm). Quantification of the proportion of untranslated mRNA granules is shown. n = 30 cells per group. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. Source data are provided as a Source Data file.

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