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Fused has evolved divergent roles in vertebrate Hedgehog signalling and motile ciliogenesis

Nature volume 459, pages 98–102 (2009)Cite this article

Abstract

Hedgehog (Hh) signalling is essential for several aspects of embryogenesis1,2. In Drosophila, Hh transduction is mediated by a cytoplasmic signalling complex3,4,5 that includes the putative serine-threonine kinase Fused (Fu) and the kinesin Costal 2 (Cos2, also known as Cos), yet Fu does not have a conserved role in Hh signalling in mammals6,7. Mouse Fu (also known as Stk36) mutants are viable and seem to respond normally to Hh signalling. Here we show that mouse Fu is essential for construction of the central pair apparatus of motile, 9+2 cilia and offers a new model of human primary ciliary dyskinesia. We found that mouse Fu physically interacts with Kif27, a mammalian Cos2 orthologue8, and linked Fu to known structural components of the central pair apparatus, providing evidence for the first regulatory component involved in central pair construction. We also demonstrated that zebrafish Fu is required both for Hh signalling and cilia biogenesis in Kupffer’s vesicle. Mouse Fu rescued both Hh-dependent and -independent defects in zebrafish. Our results delineate a new pathway for central pair apparatus assembly, identify common regulators of Hh signalling and motile ciliogenesis, and provide insights into the evolution of the Hh cascade.

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Figure 1: Mouse Fu is required for central pair apparatus construction.
Figure 2: Mouse Fu is capable of rescuing Hh-related phenotypes in zebrafish fu morphants.
Figure 3: Zebrafish fu has a Hh-independent role in left–right asymmetry and generation of 9+2 cilia.
Figure 4: Mouse Fu interacts with the central pair protein Spag16 and the Cos2 orthologue Kif27.

References

  1. McMahon, A. P., Ingham, P. W. & Tabin, C. J. Developmental roles and clinical significance of hedgehog signaling. Curr. Top. Dev. Biol. 53, 1–114 (2003)

    Article  CAS  PubMed  Google Scholar 

  2. Huangfu, D. & Anderson, K. V. Signaling from Smo to Ci/Gli: conservation and divergence of Hedgehog pathways from Drosophila to vertebrates. Development 133, 3–14 (2006)

    Article  CAS  PubMed  Google Scholar 

  3. Sisson, J. C., Ho, K. S., Suyama, K. & Scott, M. P. Costal2, a novel kinesin-related protein in the Hedgehog signaling pathway. Cell 90, 235–245 (1997)

    Article  CAS  PubMed  Google Scholar 

  4. Robbins, D. J. et al. Hedgehog elicits signal transduction by means of a large complex containing the kinesin-related protein costal2. Cell 90, 225–234 (1997)

    Article  CAS  PubMed  Google Scholar 

  5. Lum, L. et al. Hedgehog signal transduction via Smoothened association with a cytoplasmic complex scaffolded by the atypical kinesin, Costal-2. Mol. Cell 12, 1261–1274 (2003)

    Article  CAS  PubMed  Google Scholar 

  6. Chen, M. H., Gao, N., Kawakami, T. & Chuang, P. T. Mice deficient in the fused homolog do not exhibit phenotypes indicative of perturbed hedgehog signaling during embryonic development. Mol. Cell. Biol. 25, 7042–7053 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Merchant, M. et al. Loss of the serine/threonine kinase fused results in postnatal growth defects and lethality due to progressive hydrocephalus. Mol. Cell. Biol. 25, 7054–7068 (2005)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Katoh, Y. & Katoh, M. KIF27 is one of orthologs for Drosophila Costal-2. Int. J. Oncol. 25, 1875–1880 (2004)

    CAS  PubMed  Google Scholar 

  9. Davenport, J. R. & Yoder, B. K. An incredible decade for the primary cilium: a look at a once-forgotten organelle. Am. J. Physiol. Renal Physiol. 289, F1159–F1169 (2005)

    Article  CAS  PubMed  Google Scholar 

  10. Davis, E. E., Brueckner, M. & Katsanis, N. The emerging complexity of the vertebrate cilium: new functional roles for an ancient organelle. Dev. Cell 11, 9–19 (2006)

    Article  CAS  PubMed  Google Scholar 

  11. Afzelius, B. A. Cilia-related diseases. J. Pathol. 204, 470–477 (2004)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Marshall, W. F. & Kintner, C. Cilia orientation and the fluid mechanics of development. Curr. Opin. Cell Biol. 20, 48–52 (2008)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Zariwala, M. A., Knowles, M. R. & Omran, H. Genetic defects in ciliary structure and function. Annu. Rev. Physiol. 69, 423–450 (2007)

    Article  CAS  PubMed  Google Scholar 

  14. McKean, P. G., Baines, A., Vaughan, S. & Gull, K. γ-Tubulin functions in the nucleation of a discrete subset of microtubules in the eukaryotic flagellum. Curr. Biol. 13, 598–602 (2003)

    Article  CAS  PubMed  Google Scholar 

  15. Chilvers, M. A., Rutman, A. & O’Callaghan, C. Ciliary beat pattern is associated with specific ultrastructural defects in primary ciliary dyskinesia. J. Allergy Clin. Immunol. 112, 518–524 (2003)

    Article  PubMed  PubMed Central  Google Scholar 

  16. Yang, X., Dillon, R. H. & Fauci, L. J. An integrative computational model of multiciliary beating. Bull. Math. Biol. 70, 1192–1215 (2008)

    Article  MathSciNet  PubMed  Google Scholar 

  17. Frisch, D. & Farbman, A. I. Development of order during ciliogenesis. Anat. Rec. 162, 221–232 (1968)

    Article  CAS  PubMed  Google Scholar 

  18. Mitchell, B., Jacobs, R., Li, J., Chien, S. & Kintner, C. A positive feedback mechanism governs the polarity and motion of motile cilia. Nature 447, 97–101 (2007)

    Article  ADS  CAS  PubMed  Google Scholar 

  19. Wolff, C., Roy, S. & Ingham, P. W. Multiple muscle cell identities induced by distinct levels and timing of hedgehog activity in the zebrafish embryo. Curr. Biol. 13, 1169–1181 (2003)

    Article  CAS  PubMed  Google Scholar 

  20. Chen, W., Burgess, S. & Hopkins, N. Analysis of the zebrafish smoothened mutant reveals conserved and divergent functions of hedgehog activity. Development 128, 2385–2396 (2001)

    CAS  PubMed  Google Scholar 

  21. Oh, S. A. et al. A divergent cellular role for the FUSED kinase family in the plant-specific cytokinetic phragmoplast. Curr. Biol. 15, 2107–2111 (2005)

    Article  CAS  PubMed  Google Scholar 

  22. Kramer-Zucker, A. G. et al. Cilia-driven fluid flow in the zebrafish pronephros, brain and Kupffer’s vesicle is required for normal organogenesis. Development 132, 1907–1921 (2005)

    Article  CAS  PubMed  Google Scholar 

  23. Shu, X. et al. Na,K-ATPase α2 and Ncx4a regulate zebrafish left-right patterning. Development 134, 1921–1930 (2007)

    Article  CAS  PubMed  Google Scholar 

  24. Neilson, L. I. et al. cDNA cloning and characterization of a human sperm antigen (SPAG6) with homology to the product of the Chlamydomonas PF16 locus. Genomics 60, 272–280 (1999)

    Article  CAS  PubMed  Google Scholar 

  25. Sapiro, R. et al. Sperm antigen 6 is the murine homologue of the Chlamydomonas reinhardtii central apparatus protein encoded by the PF16 locus. Biol. Reprod. 62, 511–518 (2000)

    Article  CAS  PubMed  Google Scholar 

  26. Zhang, Z. et al. A sperm-associated WD repeat protein orthologous to Chlamydomonas PF20 associates with Spag6, the mammalian orthologue of Chlamydomonas PF16. Mol. Cell. Biol. 22, 7993–8004 (2002)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Smith, E. F. & Lefebvre, P. A. PF20 gene product contains WD repeats and localizes to the intermicrotubule bridges in Chlamydomonas flagella. Mol. Biol. Cell 8, 455–467 (1997)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Dawe, H. R., Farr, H. & Gull, K. Centriole/basal body morphogenesis and migration during ciliogenesis in animal cells. J. Cell Sci. 120, 7–15 (2007)

    Article  CAS  PubMed  Google Scholar 

  29. Tay, S. Y., Ingham, P. W. & Roy, S. A homologue of the Drosophila kinesin-like protein Costal2 regulates Hedgehog signal transduction in the vertebrate embryo. Development 132, 625–634 (2005)

    Article  CAS  PubMed  Google Scholar 

  30. Varjosalo, M., Li, S. P. & Taipale, J. Divergence of hedgehog signal transduction mechanism between Drosophila and mammals. Dev. Cell 10, 177–186 (2006)

    Article  CAS  PubMed  Google Scholar 

  31. Westerfield, M. The Zebrafish Book (Univ. Oregon Press, 1995)

    Google Scholar 

  32. Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R. Manipulating the Mouse Embryo: A Laboratory Manual 3rd edn (Cold Spring Harbour Laboratory Press, 2003)

    Google Scholar 

  33. Sambrook, J. & Russell, D. W. Molecular Cloning: A Laboratory Manual (Cold Spring Harbour Laboratory Press, 2001)

    Google Scholar 

  34. Eisen, J. S. & Smith, J. C. Controlling morpholino experiments: don’t stop making antisense. Development 135, 1735–1743 (2008)

    Article  CAS  PubMed  Google Scholar 

  35. Chen, J. N. & Fishman, M. C. Zebrafish tinman homolog demarcates the heart field and initiates myocardial differentiation. Development 122, 3809–3816 (1996)

    CAS  PubMed  Google Scholar 

  36. You, Y., Richer, E. J., Huang, T. & Brody, S. L. Growth and differentiation of mouse tracheal epithelial cells: selection of a proliferative population. Am. J. Physiol. Lung Cell. Mol. Physiol. 283, L1315–L1321 (2002)

    Article  CAS  PubMed  Google Scholar 

  37. Vladar, E. K. & Stearns, T. Molecular characterization of centriole assembly in ciliated epithelial cells. J. Cell Biol. 178, 31–42 (2007)

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank H. Bourne, C. C. Hui, Z. Zhang, J. Strauss III and W. Hwang for constructs, antibodies and sharing of unpublished results; R. Harland and T. Mikawa for X. tropicalis and chicken tissue; K. Thorn and S. Dandekar for assistance with microscopy and ciliary beat frequency analysis; M.-L. Cheong and Y. Nozawa for technical assistance; and D. Casso, S. Coughlin, T. Kornberg, W. Marshall, T. Mikawa, K. Wemmer and members of the Chen and Chuang laboratories for discussion and critical reading of the manuscript. Some data for this study were acquired at the Nikon Imaging Center at UCSF/QB3. This work was supported by grants from the National Institutes of Health to J.-N.C. and P.-T.C., and a Career Investigator Award from the American Lung Association to P.-T.C.

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Author notes

  1. Jehn-Hsiahn Yang

    Present address: Present address: Department of Obstetrics and Gynecology, College of Medicine and the Hospital, National Taiwan University, Taipei, Taiwan.,

  2. Christopher W. Wilson and Catherine T. Nguyen: These authors contributed equally to this work.

Authors and Affiliations

  1. Cardiovascular Research Institute, University of California, San Francisco, California 94158, USA ,

    Christopher W. Wilson, Miao-Hsueh Chen, Jehn-Hsiahn Yang, Rhodora Gacayan & Pao-Tien Chuang

  2. Department of Molecular, Cell and Developmental Biology, University of California, Los Angeles, California 90095, USA,

    Catherine T. Nguyen, Jie Huang & Jau-Nian Chen

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Correspondence to Pao-Tien Chuang.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-12 with Legends and Supplementary References. (PDF 16752 kb)

Supplementary Movie 1

This movie shows tracheal flow (visualized using Fluospheres at 40x magnification) in a wild-type mouse tracheal explant (from postnatal day (p) 14). Movie was acquired at 26 frames per second (fps) and is played back at the same speed. Left is proximal and right is distal. (MOV 1308 kb)

Supplementary Movie 2

This movie shows tracheal flow (visualized and acquired as per Supplementary Movie 1) in a Fu-/- tracheal explant. In contrast to the wild-type trachea, Fu-/- mutants do not generate a coordinated, directional flow. (MOV 937 kb)

Supplementary Movie 3

This movie shows a lateral view of motile, 9+2 cilia in a wild-type mouse tracheal explant (p14). Movie was acquired using 90x magnification at 60 fps and is played back at both 60 fps and 10 fps. (MOV 4202 kb)

Supplementary Movie 4

This movie shows a lateral view of cilia in a Fu-/- tracheal explant (acquired and visualized as per Supplementary Movie 3). Cilia in the Fu-/- trachea do not beat in a coordinated manner; many are immotile or move slowly and stiffly. (MOV 3715 kb)

Supplementary Movie 5

This movie shows a top-down view of motile, 9+2 cilia tips in a wild-type tracheal explant (acquired and visualized as per Supplementary Movie 3). Movies are shown without and with traces of individual cilia paths. (MOV 448 kb)

Supplementary Movie 6

This movie shows a top-down view of cilia tips in a Fu-/- tracheal explant (acquired and visualized as per Supplementary Movie 3). Many cilia in the Fu-/- trachea have a limited range of motion, collide with their neighbours, and do not beat in a coordinated fashion. (MOV 353 kb)

Supplementary Movie 7

This movie shows a ventral view of counterclockwise fluid flow (visualized by fluorescent beads at 40x magnification) in Kupffer's vesicle (KV) from wild-type zebrafish embryos at the 8-10 somite stage. (MOV 700 kb)

Supplementary Movie 8

This movie shows fluid flow (visualized as per Supplementary Movie 7) in Kupffer's vesicle (KV) from a fu morphant at the 8-10 somite stage. Counterclockwise fluid flow is disrupted in fu morphants. (MOV 1270 kb)

Supplementary Movie 9

This movie shows fluid flow (visualized as per Supplementary Movie 7) in Kupffer's vesicle (KV) from a fu morphant co-injected with mouse Fu mRNA at the 8-10 somite stage. Counterclockwise flow is restored in fu morphants by co-injection of mouse Fu. (MOV 1142 kb)

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Wilson, C., Nguyen, C., Chen, MH. et al. Fused has evolved divergent roles in vertebrate Hedgehog signalling and motile ciliogenesis. Nature 459, 98–102 (2009). https://doi.org/10.1038/nature07883

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