Poc1 is a Basal Body Inner Junction Protein That Promotes Triplet Microtubule Integrity and Interconnections

Overview

Journal bioRxiv

Date 2023 Nov 28

PMID 38014135

Authors

Marisa D Ruehle

Sam Li

David A Agard

Chad G Pearson

Affiliations

Soon will be listed here.

Abstract

Basal bodies (BBs) are conserved eukaryotic structures that organize motile and primary cilia. The BB is comprised of nine, cylindrically arranged, triplet microtubules (TMTs) that are connected to each other by inter-TMT linkages which maintain BB structure. During ciliary beating, forces transmitted to the BB must be resisted to prevent BB disassembly. Poc1 is a conserved BB protein important for BBs to resist ciliary forces. To understand how Poc1 confers BB stability, we identified the precise position of Poc1 binding in the BB and the effect of Poc1 loss on BB structure. Poc1 binds at the TMT inner junctions, stabilizing TMTs directly. From this location, Poc1 also stabilizes inter-TMT linkages throughout the BB, including the cartwheel pinhead and the inner scaffold. Moreover, we identify a molecular response to ciliary forces via a molecular remodeling of the inner scaffold, as determined by differences in Fam161A localization. Thus, while not essential for BB assembly, Poc1 promotes BB interconnections that establish an architecture competent to resist ciliary forces.

References

Mercey O, Kostic C, Bertiaux E, Giroud A, Sadian Y, Gaboriau D . The connecting cilium inner scaffold provides a structural foundation that protects against retinal degeneration. PLoS Biol. 2022; 20(6):e3001649. PMC: 9202906. DOI: 10.1371/journal.pbio.3001649. View

Hiraki M, Nakazawa Y, Kamiya R, Hirono M . Bld10p constitutes the cartwheel-spoke tip and stabilizes the 9-fold symmetry of the centriole. Curr Biol. 2007; 17(20):1778-83. DOI: 10.1016/j.cub.2007.09.021. View

Le Guennec M, Klena N, Gambarotto D, Laporte M, Tassin A, van den Hoek H . A helical inner scaffold provides a structural basis for centriole cohesion. Sci Adv. 2020; 6(7):eaaz4137. PMC: 7021493. DOI: 10.1126/sciadv.aaz4137. View

Arslanhan M, Cengiz-Emek S, Odabasi E, Steib E, Hamel V, Guichard P . CCDC15 localizes to the centriole inner scaffold and controls centriole length and integrity. J Cell Biol. 2023; 222(12). PMC: 10630097. DOI: 10.1083/jcb.202305009. View

Jumper J, Evans R, Pritzel A, Green T, Figurnov M, Ronneberger O . Highly accurate protein structure prediction with AlphaFold. Nature. 2021; 596(7873):583-589. PMC: 8371605. DOI: 10.1038/s41586-021-03819-2. View

Frank J, Radermacher M, Penczek P, Zhu J, Li Y, Ladjadj M . SPIDER and WEB: processing and visualization of images in 3D electron microscopy and related fields. J Struct Biol. 1996; 116(1):190-9. DOI: 10.1006/jsbi.1996.0030. View

Xu C, Min J . Structure and function of WD40 domain proteins. Protein Cell. 2011; 2(3):202-14. PMC: 4875305. DOI: 10.1007/s13238-011-1018-1. View

Kitagawa D, Vakonakis I, Olieric N, Hilbert M, Keller D, Olieric V . Structural basis of the 9-fold symmetry of centrioles. Cell. 2011; 144(3):364-75. PMC: 3089914. DOI: 10.1016/j.cell.2011.01.008. View

Scheres S, Chen S . Prevention of overfitting in cryo-EM structure determination. Nat Methods. 2012; 9(9):853-4. PMC: 4912033. DOI: 10.1038/nmeth.2115. View

10.

Li S, Fernandez J, Marshall W, Agard D . Three-dimensional structure of basal body triplet revealed by electron cryo-tomography. EMBO J. 2011; 31(3):552-62. PMC: 3273388. DOI: 10.1038/emboj.2011.460. View

11.

Grandy C, Port F, Radzinski M, Singh K, Erz D, Pfeil J . Remodeling of the focal adhesion complex by hydrogen-peroxide-induced senescence. Sci Rep. 2023; 13(1):9735. PMC: 10272183. DOI: 10.1038/s41598-023-36347-2. View

12.

Yanagisawa H, Mathis G, Oda T, Hirono M, Richey E, Ishikawa H . FAP20 is an inner junction protein of doublet microtubules essential for both the planar asymmetrical waveform and stability of flagella in Chlamydomonas. Mol Biol Cell. 2014; 25(9):1472-83. PMC: 4004596. DOI: 10.1091/mbc.E13-08-0464. View

13.

Fernandez J, Li S, Agard D . Consideration of sample motion in cryo-tomography based on alignment residual interpolation. J Struct Biol. 2019; 205(3):1-6. PMC: 6420838. DOI: 10.1016/j.jsb.2019.01.005. View

14.

Beck B, Phillips J, Bartram M, Wegner J, Thoenes M, Pannes A . Mutation of POC1B in a severe syndromic retinal ciliopathy. Hum Mutat. 2014; 35(10):1153-62. PMC: 4425427. DOI: 10.1002/humu.22618. View

15.

Allen R . The morphogenesis of basal bodies and accessory structures of the cortex of the ciliated protozoan Tetrahymena pyriformis. J Cell Biol. 1969; 40(3):716-33. PMC: 2107651. DOI: 10.1083/jcb.40.3.716. View

16.

Wickstead B, Gull K . The evolution of the cytoskeleton. J Cell Biol. 2011; 194(4):513-25. PMC: 3160578. DOI: 10.1083/jcb.201102065. View

17.

Meehl J, Bayless B, Giddings Jr T, Pearson C, Winey M . Tetrahymena Poc1 ensures proper intertriplet microtubule linkages to maintain basal body integrity. Mol Biol Cell. 2016; 27(15):2394-403. PMC: 4966981. DOI: 10.1091/mbc.E16-03-0165. View

18.

Carvalho-Santos Z, Azimzadeh J, Pereira-Leal J, Bettencourt-Dias M . Evolution: Tracing the origins of centrioles, cilia, and flagella. J Cell Biol. 2011; 194(2):165-75. PMC: 3144413. DOI: 10.1083/jcb.201011152. View

19.

Bobinnec Y, Khodjakov A, Mir L, Rieder C, Edde B, Bornens M . Centriole disassembly in vivo and its effect on centrosome structure and function in vertebrate cells. J Cell Biol. 1998; 143(6):1575-89. PMC: 2132987. DOI: 10.1083/jcb.143.6.1575. View

20.

Li S, Fernandez J, Marshall W, Agard D . Electron cryo-tomography provides insight into procentriole architecture and assembly mechanism. Elife. 2019; 8. PMC: 6384029. DOI: 10.7554/eLife.43434. View