Active Transport and Diffusion Barriers Restrict Joubert Syndrome-Associated ARL13B/ARL-13 to an Inv-like Ciliary Membrane Subdomain

English version České info

Cilia are microtubule-based cell appendages, serving motility, chemo-/mechano-/photo- sensation, and developmental signaling functions. Cilia are comprised of distinct structural and functional subregions including the basal body, transition zone (TZ) and inversin (Inv) compartments, and defects in this organelle are associated with an expanding spectrum of inherited disorders including Bardet-Biedl syndrome (BBS), Meckel-Gruber Syndrome (MKS), Joubert Syndrome (JS) and Nephronophthisis (NPHP). Despite major advances in understanding ciliary trafficking pathways such as intraflagellar transport (IFT), how proteins are transported to subciliary membranes remains poorly understood. Using Caenorhabditis elegans and mammalian cells, we investigated the transport mechanisms underlying compartmentalization of JS-associated ARL13B/ARL-13, which we previously found is restricted at proximal ciliary membranes. We now show evolutionary conservation of ARL13B/ARL-13 localisation to an Inv-like subciliary membrane compartment, excluding the TZ, in many C. elegans ciliated neurons and in a subset of mammalian ciliary subtypes. Compartmentalisation of C. elegans ARL-13 requires a C-terminal RVVP motif and membrane anchoring to prevent distal cilium and nuclear targeting, respectively. Quantitative imaging in more than 20 mutants revealed differential contributions for IFT and ciliopathy modules in defining the ARL-13 compartment; IFT-A/B, IFT-dynein and BBS genes prevent ARL-13 accumulation at periciliary membranes, whereas MKS/NPHP modules additionally inhibit ARL-13 association with TZ membranes. Furthermore, in vivo FRAP analyses revealed distinct roles for IFT and MKS/NPHP genes in regulating a TZ barrier to ARL-13 diffusion, and intraciliary ARL-13 diffusion. Finally, C. elegans ARL-13 undergoes IFT-like motility and quantitative protein complex analysis of human ARL13B identified functional associations with IFT-B complexes, mapped to IFT46 and IFT74 interactions. Together, these findings reveal distinct requirements for sequence motifs, IFT and ciliopathy modules in defining an ARL-13 subciliary membrane compartment. We conclude that MKS/NPHP modules comprise a TZ barrier to ARL-13 diffusion, whereas IFT genes predominantly facilitate ARL-13 ciliary entry and/or retention via active transport mechanisms.

Vyšlo v časopise: Active Transport and Diffusion Barriers Restrict Joubert Syndrome-Associated ARL13B/ARL-13 to an Inv-like Ciliary Membrane Subdomain. PLoS Genet 9(12): e32767. doi:10.1371/journal.pgen.1003977
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003977

Souhrn

Zdroje

1. FischC, Dupuis-WilliamsP (2011) Ultrastructure of cilia and flagella - back to the future!. Biol Cell 103: 249–319.

2. MarshallWF (2008) Basal bodies platforms for building cilia. Curr Top Dev Biol 85: 1–22.

3. ReiterJF, BlacqueOE, LerouxMR (2012) The base of the cilium: roles for transition fibres and the transition zone in ciliary formation, maintenance and compartmentalization. EMBO Rep 13: 608–18.

4. ShibaD, YamaokaY, HagiwaraH, TakamatsuT, HamadaH, et al. (2009) Localization of Inv in a distinctive intraciliary compartment requires the C-terminal ninein-homolog-containing region. J Cell Sci 122: 44–54.

5. Warburton-PittS, JaureguiA, LiC, WangJ, LerouxM, et al. (2012) Ciliogenesis in Caenorhabditis elegans requires genetic interactions between ciliary middle segment localized NPHP-2 (inversin) and transition zone-associated proteins. J Cell Sci 125: 2592–3195.

6. BlacqueOE, CevikS, KaplanOI (2008) Intraflagellar transport: from molecular characterisation to mechanism. Front Biosci 13: 2633–2652.

7. IshikawaH, MarshallW (2011) Ciliogenesis: building the cell's antenna. Nature reviews Molecular cell biology 12: 222–256.

8. QinH, BurnetteDT, BaeYK, ForscherP, BarrMM, et al. (2005) Intraflagellar transport is required for the vectorial movement of TRPV channels in the ciliary membrane. Curr Biol 15: 1695–1699.

9. HuangK, DienerD, MitchellA, PazourG, WitmanG, et al. (2007) Function and dynamics of PKD2 in Chlamydomonas reinhardtii flagella. J Cell Biol 179: 501–514.

10. HaoL, TheinM, Brust-MascherI, Civelekoglu-ScholeyG, LuY, et al. (2011) Intraflagellar transport delivers tubulin isotypes to sensory cilium middle and distal segments. Nat Cell Biol 13: 790–798.

11. BlacqueOE, ReardonMJ, LiC, McCarthyJ, MahjoubMR, et al. (2004) Loss of C. elegans BBS-7 and BBS-8 protein function results in cilia defects and compromised intraflagellar transport. Genes Dev 18: 1630–1642.

12. OuG, BlacqueOE, SnowJJ, LerouxMR, ScholeyJM (2005) Functional coordination of intraflagellar transport motors. Nature 436: 583–587.

13. LechtreckK-F, JohnsonE, SakaiT, CochranD, BallifB, et al. (2009) The Chlamydomonas reinhardtii BBSome is an IFT cargo required for export of specific signaling proteins from flagella. J Cell Biol 187: 1117–1149.

14. WeiQ, ZhangY, LiY, ZhangQ, LingK, et al. (2012) The BBSome controls IFT assembly and turnaround in cilia. Nat Cell Biol 14: 950–957.

15. NachuryMV, SeeleyES, JinH (2010) Trafficking to the ciliary membrane: how to get across the periciliary diffusion barrier? Annu Rev Cell Dev Biol 26: 59–87.

16. CraigeB, TsaoCC, DienerDR, HouY, LechtreckKF, et al. (2010) CEP290 tethers flagellar transition zone microtubules to the membrane and regulates flagellar protein content. J Cell Biol 190: 927–940.

17. ChihB, LiuP, ChinnY, ChalouniC, KomuvesLG, et al. (2011) A ciliopathy complex at the transition zone protects the cilia as a privileged membrane domain. Nat Cell Biol 14: 61–72.

18. Garcia-GonzaloFR, CorbitKC, Sirerol-PiquerMS, RamaswamiG, OttoEA, et al. (2011) A transition zone complex regulates mammalian ciliogenesis and ciliary membrane composition. Nat Genet 43: 776–784.

19. WilliamsCL, LiC, KidaK, InglisPN, MohanS, et al. (2011) MKS and NPHP modules cooperate to establish basal body/transition zone membrane associations and ciliary gate function during ciliogenesis. J Cell Biol 192: 1023–1041.

20. HuangL, SzymanskaK, JensenVL, JaneckeAR, InnesAM, et al. (2011) TMEM237 Is Mutated in Individuals with a Joubert Syndrome Related Disorder and Expands the Role of the TMEM Family at the Ciliary Transition Zone. Am J Hum Genet 89: 713–730.

21. HuQ, MilenkovicL, JinH, ScottMP, NachuryMV, et al. (2010) A septin diffusion barrier at the base of the primary cilium maintains ciliary membrane protein distribution. Science 329: 436–439.

22. KimS, ShindoA, ParkT, OhE, GhoshS, et al. (2010) Planar cell polarity acts through septins to control collective cell movement and ciliogenesis. Science (New York, NY) 329: 1337–1377.

23. FanS, FoggV, WangQ, ChenXW, LiuCJ, et al. (2007) A novel Crumbs3 isoform regulates cell division and ciliogenesis via importin beta interactions. J Cell Biol 178: 387–398.

24. DishingerJF, KeeHL, JenkinsPM, FanS, HurdTW, et al. (2010) Ciliary entry of the kinesin-2 motor KIF17 is regulated by importin-beta2 and RanGTP. Nat Cell Biol 12: 703–710.

25. FanS, WhitemanEL, HurdTW, McIntyreJC, DishingerJE, et al. (2011) Induction of Ran GTP Drives Ciliogenesis. Mol Biol Cell 4539–48.

26. HurdTW, FanS, MargolisBL (2011) Localization of retinitis pigmentosa 2 to cilia is regulated by Importin beta2. J Cell Sci 124: 718–726.

27. KeeHL, DishingerJF, BlasiusTL, LiuCJ, MargolisB, et al. (2012) A size-exclusion permeability barrier and nucleoporins characterize a ciliary pore complex that regulates transport into cilia. Nat Cell Biol 14: 431–437.

28. Inglis PN, Ou G, Leroux MR, Scholey JM (2007) The sensory cilia of C. elegans. WormBook, ed The C elegans Research Community: WormBook. doi/101895/wormbook11231.

29. PazourG, San AgustinJ, FollitJ, RosenbaumJ, WitmanG (2002) Polycystin-2 localizes to kidney cilia and the ciliary level is elevated in orpk mice with polycystic kidney disease. Current biology : CB 12: 80.

30. BaeYK, QinH, KnobelKM, HuJ, RosenbaumJL, et al. (2006) General and cell-type specific mechanisms target TRPP2/PKD-2 to cilia. Development 133: 3859–3870.

31. CantagrelV, SilhavyJL, BielasSL, SwistunD, MarshSE, et al. (2008) Mutations in the cilia gene ARL13B lead to the classical form of Joubert syndrome. Am J Hum Genet 83: 170–179.

32. CasparyT, LarkinsCE, AndersonKV (2007) The graded response to Sonic Hedgehog depends on cilia architecture. Dev Cell 12: 767–778.

33. HoriY, KobayashiT, KikkoY, KontaniK, KatadaT (2008) Domain architecture of the atypical Arf-family GTPase Arl13b involved in cilia formation. Biochem Biophys Res Commun 373: 119–124.

34. DuldulaoNA, LeeS, SunZ (2009) Cilia localization is essential for in vivo functions of the Joubert syndrome protein Arl13b/Scorpion. Development 136: 4033–4042.

35. CevikS, HoriY, KaplanOI, KidaK, ToivenonT, et al. (2010) Joubert syndrome Arl13b functions at ciliary membranes and stabilizes protein transport in Caenorhabditis elegans. J Cell Biol 188: 953–969.

36. LiY, WeiQ, ZhangY, LingK, HuJ (2010) The small GTPases ARL-13 and ARL-3 coordinate intraflagellar transport and ciliogenesis. J Cell Biol 189: 1039–1051.

37. LarkinsC, AvilesG, EastM, KahnR, CasparyT (2011) Arl13b regulates ciliogenesis and the dynamic localization of Shh signaling proteins. Mol Biol Cell 22: 4694–5397.

38. HigginbothamH, EomTY, MarianiLE, BachledaA, HirtJ, et al. (2012) Arl13b in primary cilia regulates the migration and placement of interneurons in the developing cerebral cortex. Dev Cell 23: 925–938.

39. HumbertMC, WeihbrechtK, SearbyCC, LiY, PopeRM, et al. (2012) ARL13B, PDE6D, and CEP164 form a functional network for INPP5E ciliary targeting. Proc Natl Acad Sci U S A 109: 19691–6.

40. LiY, ZhangQ, WeiQ, ZhangY, LingK, et al. (2012) SUMOylation of the small GTPase ARL-13 promotes ciliary targeting of sensory receptors. J Cell Biol 199: 589–598.

41. SnowJJ, OuG, GunnarsonAL, WalkerMR, ZhouHM, et al. (2004) Two anterograde intraflagellar transport motors cooperate to build sensory cilia on C. elegans neurons. Nat Cell Biol 6: 1109–1113.

42. MazelovaJ, Astuto-GribbleL, InoueH, TamBM, SchonteichE, et al. (2009) Ciliary targeting motif VxPx directs assembly of a trafficking module through Arf4. Embo J 28: 183–192.

43. ChunDK, McEwenJM, BurbeaM, KaplanJM (2008) UNC-108/Rab2 regulates postendocytic trafficking in Caenorhabditis elegans. Mol Biol Cell 19: 2682–2695.

44. HsiaoY-C, TuzK, FerlandR (2012) Trafficking in and to the primary cilium. Cilia 1: 4.

45. OuG, KogaM, BlacqueOE, MurayamaT, OhshimaY, et al. (2007) Sensory Ciliogenesis in Caenorhabditis elegans: Assignment of IFT components into Distinct Modules Based on Transport and Phenotypic Profiles. Mol Biol Cell 18: 1554–69.

46. MukhopadhyayS, WenX, ChihB, NelsonC, LaneW, et al. (2010) TULP3 bridges the IFT-A complex and membrane phosphoinositides to promote trafficking of G protein-coupled receptors into primary cilia. Genes Dev 24: 2180–2193.

47. LechtreckK, BrownJ, SampaioJ, CraftJ, ShevchenkoA, et al. (2013) Cycling of the signaling protein phospholipase D through cilia requires the BBSome only for the export phase. J Cell Biol 201: 249–261.

48. PanF, MalmbergR, MomanyM (2007) Analysis of septins across kingdoms reveals orthology and new motifs. BMC evolutionary biology 7: 103.

49. PerkinsLA, HedgecockEM, ThomsonJN, CulottiJG (1986) Mutant sensory cilia in the nematode Caenorhabditis elegans. Dev Biol 117: 456–487.

50. GloecknerCJ, BoldtK, SchumacherA, RoepmanR, UeffingM (2007) A novel tandem affinity purification strategy for the efficient isolation and characterisation of native protein complexes. Proteomics 7: 4228–4234.

51. LuckerBF, BehalRH, QinH, SironLC, TaggartWD, et al. (2005) Characterization of the intraflagellar transport complex B core: direct interaction of the IFT81 and IFT74/72 subunits. J Biol Chem 280: 27688–27696.

52. LuckerBF, MillerMS, DziedzicSA, BlackmarrPT, ColeDG (2010) Direct interactions of intraflagellar transport complex B proteins IFT88, IFT52, and IFT46. J Biol Chem 285: 21508–21518.

53. SangL, MillerJJ, CorbitKC, GilesRH, BrauerMJ, et al. (2011) Mapping the NPHP-JBTS-MKS protein network reveals ciliopathy disease genes and pathways. Cell 145: 513–528.

54. BoldtK, MansDA, WonJ, van ReeuwijkJ, VogtA, et al. (2011) Disruption of intraflagellar protein transport in photoreceptor cilia causes Leber congenital amaurosis in humans and mice. J Clin Invest 121: 2169–2180.

55. ShibaD, ManningD, KogaH, BeierD, YokoyamaT (2010) Inv acts as a molecular anchor for Nphp3 and Nek8 in the proximal segment of primary cilia. Cytoskeleton (Hoboken, NJ) 67: 112–119.

56. FollitJ, XuF, KeadyB, PazourG (2009) Characterization of mouse IFT complex B. Cell Motil Cytoskeleton 66: 457–468.

57. FranklinJ, UlluE (2010) Biochemical analysis of PIFTC3, the Trypanosoma brucei orthologue of nematode DYF-13, reveals interactions with established and putative intraflagellar transport components. Mol Microbiol 78: 173–186.

58. BlacqueOE, PerensEA, BoroevichKA, InglisPN, LiC, et al. (2005) Functional genomics of the cilium, a sensory organelle. Curr Biol 15: 935–941.

59. ZhangQ, LiuQ, AustinC, DrummondI, PierceE (2012) Knockdown of ttc26 disrupts ciliogenesis of the photoreceptor cells and the pronephros in zebrafish. Mol Biol Cell 23: 3069–3078.

60. NakadaC, RitchieK, ObaY, NakamuraM, HottaY, et al. (2003) Accumulation of anchored proteins forms membrane diffusion barriers during neuronal polarization. Nat Cell Biol 5: 626–632.

61. SarikasA, HartmannT, PanZ-Q (2011) The cullin protein family. Genome Biol 12: 220.

62. BrennerS (1974) The genetics of Caenorhabditis elegans. Genetics 77: 71–94.

63. HobertO (2002) PCR fusion-based approach to create reporter gene constructs for expression analysis in transgenic C. elegans. Biotechniques 32: 728–730.

64. GloecknerCJ, BoldtK, UeffingM (2009) Strep/FLAG tandem affinity purification (SF-TAP) to study protein interactions. Curr Protoc Protein Sci Chapter 19: Unit19 20.

65. BendallSC, HughesC, StewartMH, DobleB, BhatiaM, et al. (2008) Prevention of amino acid conversion in SILAC experiments with embryonic stem cells. Mol Cell Proteomics 7: 1587–1597.

66. OlsenJV, de GodoyLM, LiG, MacekB, MortensenP, et al. (2005) Parts per million mass accuracy on an Orbitrap mass spectrometer via lock mass injection into a C-trap. Mol Cell Proteomics 4: 2010–2021.

67. CoxJ, MannM (2008) MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification. Nat Biotechnol 26: 1367–1372.

68. LetteboerSJ, RoepmanR (2008) Versatile screening for binary protein-protein interactions by yeast two-hybrid mating. Methods Mol Biol 484: 145–159.