Requirement of the Cep57-Cep63 Interaction for Proper Cep152 Recruitment and Centriole Duplication

ABSTRACT

Cep57 has been characterized as a component of a pericentriolar complex containing Cep63 and Cep152. Interestingly, Cep63 and Cep152 self-assemble into a pericentriolar cylindrical architecture, and this event is critical for the orderly recruitment of Plk4, a key regulator of centriole duplication. However, the way in which Cep57 interacts with the Cep63-Cep152 complex and contributes to the structure and function of Cep63-Cep152 self-assembly remains unknown. We demonstrate that Cep57 interacts with Cep63 through N-terminal motifs and associates with Cep152 via Cep63. Three-dimensional structured illumination microscopy (3D-SIM) analyses suggested that the Cep57-Cep63-Cep152 complex is concentrically arranged around a centriole in a Cep57-in and Cep152-out manner. Cep57 mutant cells defective in Cep63 binding exhibited improper Cep63 and Cep152 localization and impaired Sas6 recruitment for procentriole assembly, proving the significance of the Cep57-Cep63 interaction. Intriguingly, Cep63 fused to a microtubule (MT)-binding domain of Cep57 functioned in concert with Cep152 to assemble around stabilized MTs in vitro. Thus, Cep57 plays a key role in architecting the Cep63-Cep152 assembly around centriolar MTs and promoting centriole biogenesis. This study may offer a platform to investigate how the organization and function of the pericentriolar architecture are altered by disease-associated mutations found in the Cep57-Cep63-Cep152 complex.

KEYWORDS:

• Cep57
• Cep63
• Cep152
• PCM
• centriole biogenesis

SUPPLEMENTAL MATERIAL

Supplemental material is available online only.

ACKNOWLEDGMENTS

We are grateful to Michael Kruhlak and Langston Lim for assistance with time-lapse microscopy, Kunio Nagashima for providing technical service with electron microscopy samples, Thorkell Andresson for carrying out the second phase of the mass spectrometry database search using Proteome Discoverer, Yanling Liu for generating 3D surface-rendered movies, and Kunsoo Rhee at Seoul National University, South Korea, for reagents.

This research was supported by the Intramural Research Program of the National Cancer Institute, National Institutes of Health (K.S.L.); an NST grant (CAP-16-03-KRIBB) of South Korea (J.K.B.); a National Research Foundation grant (2019M3E5D6063955) funded by the Ministry of Science and ICT of South Korea (B.K.); and a Korean Biomedical Scientist fellowship (KGS1001911) from the KRIBB Research Initiative Program (J.I.A.).