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Super-resolution microscopy reveals that disruption of ciliary transition-zone architecture causes Joubert syndrome

An Erratum to this article was published on 31 October 2017

This article has been updated


Ciliopathies, including nephronophthisis (NPHP), Meckel syndrome (MKS) and Joubert syndrome (JBTS), can be caused by mutations affecting components of the transition zone, a domain near the base of the cilium that controls the protein composition of its membrane. We defined the three-dimensional arrangement of key proteins in the transition zone using two-colour stochastic optical reconstruction microscopy (STORM). NPHP and MKS complex components form nested rings comprised of nine-fold doublets. JBTS-associated mutations in RPGRIP1L or TCTN2 displace certain transition-zone proteins. Diverse ciliary proteins accumulate at the transition zone in wild-type cells, suggesting that the transition zone is a waypoint for proteins entering and exiting the cilium. JBTS-associated mutations in RPGRIP1L disrupt SMO accumulation at the transition zone and the ciliary localization of SMO. We propose that the disruption of transition-zone architecture in JBTS leads to a failure of SMO to accumulate at the transition zone and cilium, disrupting developmental signalling in JBTS.

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Figure 1: MKS and NPHP complexes form rings of discrete puncta.
Figure 2: MKS and NPHP complex rings are comprised of doublets.
Figure 3: MKS and NPHP complexes occupy the same proximodistal position in the transition zone, where SMO docks following Hedgehog pathway activation.
Figure 4: MKS and NPHP complexes are displaced from the transition zone in a Joubert syndrome patient with mutations in TCTN2.
Figure 5: MKS and NPHP complexes are displaced from the transition zone in Joubert syndrome patients with mutations in RPGRIP1L.
Figure 6: Transition-zone rings formed by MKS and NPHP complexes are disrupted in a Joubert syndrome patient with mutations in RPGRIP1L.

Change history

  • 25 September 2017

    In the original version of this Article, the name of author Galo Garcia III was coded wrongly, resulting in it being incorrect when exported to citation databases. This has now been corrected, though no visible changes will be apparent.


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Structured illumination microscopy was performed on a Nikon N-SIM system in the UCSF Nikon Imaging Center. We thank H. Liu and J. Schnitzbauer for help setting up the STORM system and J. Schnitzbauer in developing the analysis algorithms. We thank V. Herranz-Pérez and J. M. Garcia-Verdugo for providing electron micrographs. This project is supported by the NIH Director’s New Innovator Award (DP2OD008479) to X.S., R.M. and B.H. and by grants from the NIH (AR054396 and GM095941 to J.F.R., F32GM109714 to G.G.III, and U54HD083091 sub-project 6849 to D.D.) and the Burroughs Wellcome Fund and the Packard Foundation to G.G.III and J.F.R. B.H. is a Chan Zuckerberg Biohub investigator.

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X.S., G.G.III, B.H. and J.F.R. designed the experiments and wrote the manuscript; X.S. and R.M. built the STORM microscope; G.G.III generated the samples; X.S. and G.G.III performed the STORM imaging experiments; and X.S. analysed the STORM data. G.G.III performed the SIM imaging experiments and analysed the SIM data. J.C.V.D.W. and D.D. collected and genotyped the human fibroblasts and provided feedback on the manuscript. G.J.P. generated the α-NPHP1 antibody.

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Correspondence to Bo Huang or Jeremy F. Reiter.

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The authors declare no competing financial interests.

Integrated supplementary information

Supplementary Figure 1

Single color 3D STORM images of SMO in (A) Ptch1−/− and (B) Wild type MEFs. The STORM images are overlaid with conventional wide-field fluorescence images. Scale bars: 500 nm.

Supplementary Figure 2 RPGRIP1L domain architecture and JBTS patient mutations.

RPGRIP1L contains seven coiled-coil segments and 3 C2 domains, including an RPGR-interacting C2 domain (RID). Patient 15-4 carries RPGRIP1L(Thr615Pro); 4-4 and 4-3 carry RPGRIP1L(Ser659Pro). Each JBTS-affected patient also carries a nonsense mutation affecting the C2-N domain or the C2-C domain. Patient 15-4 carries RPGRIP1L(Gln684X); patients 4-4 and 4-3 carry RPGRIP1L(Arg805X).

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Shi, X., Garcia, G., Van De Weghe, J. et al. Super-resolution microscopy reveals that disruption of ciliary transition-zone architecture causes Joubert syndrome. Nat Cell Biol 19, 1178–1188 (2017).

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