Molecular Organization of the Distal Tip of Vertebrate Motile Cilia

Overview

Journal bioRxiv

Date 2025 Mar 3

PMID 40027778

Authors

Juyeon Hong

Chanjae Lee

Ophelia Papoulas

Jiehong Pan

Maki Takagishi

Nadia Manzi

Daniel J Dickinson

Amjad Horani

Steven L Brody

Edward Marcotte

Tae Joo Park

John B Wallingford

Affiliations

Soon will be listed here.

Abstract

The beating of cilia on multi-ciliated cells (MCCs) is essential for normal development and homeostasis in animals. Unlike basal bodies or axonemes, the distal tips of MCC cilia remain poorly defined. Here, we characterize the molecular organization of the distal tip of vertebrate MCC cilia, revealing two distinct domains occupied by distinct protein constituents. Using frog, mouse, and human MCCs, we find that two largely uncharacterized proteins, Ccdc78 and Ccdc33 occupy a specialized region at the extreme distal tip, and these are required for the normal organization of other tip proteins, including Spef1, Cep104, and Eb3. Ccdc78 and Cccdc33 are also independently required for normal length regulation of MCC cilia. Mechanistically, Ccdc78 and Ccdc33 display robust microtubule-bundling activity both and . Thus, we reveal that two previously undefined proteins form a key module for organizing and stabilizing the distal tip of motile cilia in vertebrate MCC. We propose that these proteins represent potential disease loci for motile ciliopathies.

References

Boon M, Wallmeier J, Ma L, Loges N, Jaspers M, Olbrich H . MCIDAS mutations result in a mucociliary clearance disorder with reduced generation of multiple motile cilia. Nat Commun. 2014; 5:4418. DOI: 10.1038/ncomms5418. View

Chung M, Kwon T, Tu F, Brooks E, Gupta R, Meyer M . Coordinated genomic control of ciliogenesis and cell movement by RFX2. Elife. 2014; 3:e01439. PMC: 3889689. DOI: 10.7554/eLife.01439. View

Horani A, Ustione A, Huang T, Firth A, Pan J, Gunsten S . Establishment of the early cilia preassembly protein complex during motile ciliogenesis. Proc Natl Acad Sci U S A. 2018; 115(6):E1221-E1228. PMC: 5819421. DOI: 10.1073/pnas.1715915115. View

Legal T, Parra M, Tong M, Black C, Joachimiak E, Valente-Paterno M . CEP104/FAP256 and associated cap complex maintain stability of the ciliary tip. J Cell Biol. 2023; 222(11). PMC: 10522465. DOI: 10.1083/jcb.202301129. View

Tu F, Sedzinski J, Ma Y, Marcotte E, Wallingford J . Protein localization screening reveals novel regulators of multiciliated cell development and function. J Cell Sci. 2017; 131(3). PMC: 5826043. DOI: 10.1242/jcs.206565. View

DENTLER W, LeCluyse E . Microtubule capping structures at the tips of tracheal cilia: evidence for their firm attachment during ciliary bend formation and the restriction of microtubule sliding. Cell Motil. 1982; 2(6):549-72. DOI: 10.1002/cm.970020605. View

Fliegauf M, Benzing T, Omran H . When cilia go bad: cilia defects and ciliopathies. Nat Rev Mol Cell Biol. 2007; 8(11):880-93. DOI: 10.1038/nrm2278. View

Gray R, Abitua P, Wlodarczyk B, Szabo-Rogers H, Blanchard O, Lee I . The planar cell polarity effector Fuz is essential for targeted membrane trafficking, ciliogenesis and mouse embryonic development. Nat Cell Biol. 2009; 11(10):1225-32. PMC: 2755648. DOI: 10.1038/ncb1966. View

Frikstad K, Molinari E, Thoresen M, Ramsbottom S, Hughes F, Letteboer S . A CEP104-CSPP1 Complex Is Required for Formation of Primary Cilia Competent in Hedgehog Signaling. Cell Rep. 2019; 28(7):1907-1922.e6. PMC: 6702141. DOI: 10.1016/j.celrep.2019.07.025. View

10.

Reynolds M, Phetruen T, Fisher R, Chen K, Pentecost B, Gomez G . The Developmental Process of the Growing Motile Ciliary Tip Region. Sci Rep. 2018; 8(1):7977. PMC: 5964098. DOI: 10.1038/s41598-018-26111-2. View

11.

Nager A, Goldstein J, Herranz-Perez V, Portran D, Ye F, Garcia-Verdugo J . An Actin Network Dispatches Ciliary GPCRs into Extracellular Vesicles to Modulate Signaling. Cell. 2016; 168(1-2):252-263.e14. PMC: 5235987. DOI: 10.1016/j.cell.2016.11.036. View

12.

Van De Weghe J, Rusterholz T, Latour B, Grout M, Aldinger K, Shaheen R . Mutations in ARMC9, which Encodes a Basal Body Protein, Cause Joubert Syndrome in Humans and Ciliopathy Phenotypes in Zebrafish. Am J Hum Genet. 2017; 101(1):23-36. PMC: 5501774. DOI: 10.1016/j.ajhg.2017.05.010. View

13.

Jessus C, Thibier C, Ozon R . Identification of microtubule-associated proteins (MAPs) in Xenopus oocyte. FEBS Lett. 1985; 192(1):135-40. DOI: 10.1016/0014-5793(85)80059-4. View

14.

Wallmeier J, Nielsen K, Kuehni C, Lucas J, Leigh M, Zariwala M . Motile ciliopathies. Nat Rev Dis Primers. 2020; 6(1):77. DOI: 10.1038/s41572-020-0209-6. View

15.

Klos Dehring D, Vladar E, Werner M, Mitchell J, Hwang P, Mitchell B . Deuterosome-mediated centriole biogenesis. Dev Cell. 2013; 27(1):103-12. PMC: 3816757. DOI: 10.1016/j.devcel.2013.08.021. View

16.

Latour B, Van De Weghe J, Rusterholz T, Letteboer S, Gomez A, Shaheen R . Dysfunction of the ciliary ARMC9/TOGARAM1 protein module causes Joubert syndrome. J Clin Invest. 2020; 130(8):4423-4439. PMC: 7410078. DOI: 10.1172/JCI131656. View

17.

Zheng J, Liu H, Zhu L, Chen Y, Zhao H, Zhang W . Microtubule-bundling protein Spef1 enables mammalian ciliary central apparatus formation. J Mol Cell Biol. 2018; 11(1):67-77. DOI: 10.1093/jmcb/mjy014. View

18.

Kaczmarek K, Niedzialkowska E, Studencka M, Schulz Y, Grzmil P . Ccdc33, a predominantly testis-expressed gene, encodes a putative peroxisomal protein. Cytogenet Genome Res. 2010; 126(3):243-52. DOI: 10.1159/000251961. View

19.

Milana E, Zhang R, Vetrano M, Peerlinck S, De Volder M, Onck P . Metachronal patterns in artificial cilia for low Reynolds number fluid propulsion. Sci Adv. 2020; 6(49). PMC: 7821886. DOI: 10.1126/sciadv.abd2508. View

20.

Chien A, Shih S, Bower R, Tritschler D, Porter M, Yildiz A . Dynamics of the IFT machinery at the ciliary tip. Elife. 2017; 6. PMC: 5662288. DOI: 10.7554/eLife.28606. View