Ferreira, David C.Carmona, BrunoNolasco, SofiaMarinho, H. SusanaSoares, Helena2020-06-112020-06-112019-11Ferreira DC, Carmona B, Nolasco S, Marinho HS, Soares H. The interplay between KIAA0753 and TBCCD1 in the control of ciliogenesis and the cell cytoskeleton architecture. In: 23rd Annual Meeting da Sociedade Portuguesa de Genética Humana, Fundação Bissaya Barreto, Bencanta (Coimbra), 14 a 16 de novembro de 2019.http://hdl.handle.net/10400.21/11822Project IPL/2019/MOONOFCILI/ESTeSLAims/Context: Cilia are slender protuberances found in eukaryotic cells, consisting of a microtubule (MT)-based ciliary axoneme, which can confer motility and sensory functions to the cells. These organelles have a basal body, which can be derived from the centrosome and nucleates the ciliary axoneme. Centriolar satellites are protein granules located around the centrosome that play an essential role in centrosome and primary cilium assembly. Mutations in genes encoding centrosome and/or centriolar satellite components lead to various human disorders, such as ciliopathies. Previous work from our group characterized the interactome of a new centrosomal TBCC domain-containing human protein (TBCCD1). Included among the identified proteins were several well-known proteins encoded by ciliopathy genes, e.g., centrosomal and centriolar satellites protein KIAA0753 (also known as OFIP and Moonraker). KIAA0753 is mutated in several ciliopathic syndromes, e.g., Joubert syndrome. Methods: Immunofluorescent microscopy and Western blot were used to study the levels and cellular localization of components of centriolar satellites, cytoskeleton, and cilia, in human RPE1 cells overexpressing TBCCD1. Similarly, RPE1 cells depleted of either TBCCD1 or KIAA0753 were analyzed. Results: In RPE-1 cells, both the knockdown and overexpression of TBCCD1 affect the levels of KIAA0753, as well as its localization around the centrosome. This suggests that TBCCD1 plays a role in the recruitment of KIAA0753 to the centrosome. Regarding the levels of KIAA0753, we show that its depletion affects centriole connection with consequences on the organization of the MT network. This may compromise cell polarity, cell migration, and cilia assembly. Furthermore, we show that KIAA0753 localizes in the basal body and along the axoneme of cilia, which points to a role in ciliogenesis. Conclusions: Our results support a new function for KIAA0753 in centrosome structure maintenance, as well as a new functional interaction between TBCCD1 and KIAA0753, suggesting a new pathway in ciliopathy development.engTBCCD1KIAA0753CiliaCentrosomeIPL/2019/MOONOFCILI/ESTeSLconference object