Functional characterization of the SAS-4-related protein CPAP in centrosome biology of human cells

Abstract

The centrosome is an organelle that resides at the center of most animal cells and comprises two microtubule-based centriole cylinders surrounded by pericentriolar material (PCM). The centrosome plays a fundamental role for nucleating and organizing a radial array of cytoplasmic microtubules during interphase and promoting the assembly of the mitotic spindle during mitosis. In proliferating cells, the centrosome duplicates once per cell cycle, a process that involves notably the formation of one procentriole at the base of each centriole. The procentriole then grows until it reaches the same size of its associated centriole. The formation of supernumerary centrosomes causes severe problems, since this can lead to multipolar spindle formation, chromosome missegregation and genomic instability, which are hallmarks of cancer cells. The mechanisms regulating centrosome duplication are still incompletely understood. We have studied the role of the centrosomal P4.1-associated protein (CPAP) in human cells. We found that this protein is required for efficient growth of cytoplasmic microtubules from centrosomes as well as for centrosome duplication. There, CPAP is required at an early step during procentriole assembly, after the incorporation of HsSAS-6 but before that of Centrin. Importantly, we found also that the overexpression of CPAP leads to abnormal elongation of procentrioles and centrioles, indicating that the levels of CPAP determine centriole length. Excess CPAP levels furthermore lead to the formation of multiple procentrioles along overly elongated centrioles, multipolar spindle formation and cell division errors. Therefore, proper regulation of proteins such as CPAP that set centriole length contributes to ensure genome integrity. Overall, we gained important insights into the functions of CPAP at the centrosome and identified a novel control mechanism of centriole length. This will be relevant for a better understanding of how centrosomes function in the progression of cancer and other diseases.