CFAP53 regulates mammalian cilia-type motility patterns through differential localization and recruitment of axonemal dynein components

Abstract

Motile cilia can beat with distinct patterns, but how motility variations are regulated remain obscure. Here, we have studied the role of the coiled-coil protein CFAP53 in the motility of different cilia-types in the mouse. While node (9+0) cilia of Cfap53 mutants were immotile, tracheal and ependymal (9+2) cilia retained motility, albeit with an altered beat pattern. In node cilia, CFAP53 mainly localized at the base (centriolar satellites), whereas it was also present along the entire axoneme in tracheal cilia. CFAP53 associated tightly with microtubules and interacted with axonemal dyneins and TTC25, a dynein docking complex component. TTC25 and outer dynein arms (ODAs) were lost from node cilia, but were largely maintained in tracheal cilia of Cfap53-/- mice. Thus, CFAP53 at the base of node cilia facilitates axonemal transport of TTC25 and dyneins, while axonemal CFAP53 in 9+2 cilia stabilizes dynein binding to microtubules. Our study establishes how differential localization and function of CFAP53 contributes to the unique motion patterns of two important mammalian cilia-types.

De novo identification of mammalian ciliary motility proteins using cryo-EM

“…Both paralogs make conserved interactions with the 24-nm ODA-DC through similar sequences in their C terminus but bind different sequences of the 48-nm filamentous MIP, CFAP53, through their diverged N termini ( Figure 5 ). The observation that CFAP53 mediates the crosstalk between MIPs and the ODA-DC explains why Cfap53 -deficient zebrafish and mice have cilia motility defects because of lack of ODAs ( Ide et al, 2020 ; Narasimhan et al, 2015 ) and why mutations in human CFAP53 cause laterality abnormalities ( Narasimhan et al, 2015 ; Noël et al, 2016 ; Perles et al, 2012 ). Using gene editing, we show that loss of pierce1 and pierce2 in zebrafish and mice affects rotating cilia more than planar beating cilia, which is consistent with previous studies showing that MIP defects mainly cause laterality abnormalities ( Table S3 ).…”

“…Third, what unites the MIP architecture and the exterior ODAs, and is this interconnectivity important for ciliary motility and function? In zebrafish and mice, genetic ablation of the suspected MIPs CFAP53 ( Ide et al, 2020 ; Narasimhan et al, 2015 ; Noël et al, 2016 ) and MNS1 ( Zhou et al, 2012 ) causes loss of ODAs from axonemes and disrupts ciliary motility. Despite evidence showing that MIPs are important for the physiologic positioning of ODAs, the molecular mechanism by which this is achieved is unknown.…”

Ciliary dynein arms: Cytoplasmic preassembly, intraflagellar transport, and axonemal docking

“…Nonetheless, the exact mechanism by which these proteins function has remained unknown. For example, CFAP53/CCDC11 was found to localize along the entire axoneme in zebrafish, mouse, and human “9 + 2” cilia, and immunoprecipitation studies have demonstrated that it can associate with the ODA‐DC component TTC25 (Ide et al, 2020). In view of this, it has been proposed that axonemal CFAP53/CCDC11 may help stabilize ODA binding on the microtubules.…”

Essential Role of CFAP53 in Sperm Flagellum Biogenesis

“…Previous studies have shown that CFAP53 is located at the basal body and on centriolar satellites in retinal pigment epithelial cells and on the ciliary axonemes in zebrafish kidneys and human respiratory cells (Narasimhan et al, 2015;Silva et al, 2016). In mice, CFAP53 is located at the base of the nodal cilia and the tracheal cilia, as well as along the axonemes of the tracheal cilia (Ide et al, 2020). Depletion of CFAP53 disrupts the subcellular organization FIGURE 5 | Localization of CFAP53 in developing germ cells.…”

“…The cilia and flagella associated protein (CFAP) family, such as CFAP58, CFAP61, CFAP69, CFAP65, CFAP43, CFAP44, CFAP70, and CFAP251, is associated with flagellum biogenesis and morphogenesis (Tang et al, 2017;Dong et al, 2018;Beurois et al, 2019;He et al, 2020;Huang et al, 2020;Li et al, 2020). Previous studies have indicated that the functional role of CFAP53 (also named the coiledcoil domain containing protein CCDC11) is involved in the biogenesis and motility of motile cilia (Perles et al, 2012;Narasimhan et al, 2015;Noël et al, 2016;Silva et al, 2016), and CFAP53 is localized not only to the base of the nodal cilia, but also along the axoneme of the tracheal cilia (Ide et al, 2020). However, the exact localization and function of CFAP53 during spermiogenesis is still poorly understood.…”