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Coilin-dependent snRNP assembly is essential for zebrafish embryogenesis

Nature Structural & Molecular Biology volume 17, pages 403–409 (2010)Cite this article

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Abstract

Spliceosomal small nuclear ribonucleoproteins (snRNPs), comprised of small nuclear RNAs (snRNAs) in complex with snRNP-specific proteins, are essential for pre-mRNA splicing. Coilin is not a snRNP protein but concentrates snRNPs and their assembly intermediates in Cajal bodies (CBs). Here we show that depletion of coilin in zebrafish embryos leads to CB dispersal, deficits in snRNP biogenesis and expression of spliced mRNA, as well as reduced cell proliferation followed by developmental arrest. Notably, injection of purified mature human snRNPs restored mRNA expression and viability. snRNAs were necessary but not sufficient for rescue, showing that only assembled snRNPs can bypass the requirement for coilin. Thus, coilin's essential function in embryos is to promote macromolecular assembly of snRNPs, likely by concentrating snRNP components in CBs to overcome rate-limiting assembly steps.

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Figure 1: Coilin is essential for embryogenesis.
Figure 2: Coilin is essential for Cajal body integrity.
Figure 3: U4 snRNA localization in living embryos reveals timing of CB dispersal upon coilin depletion.
Figure 4: Coilin morphants show defects in snRNP assembly and expression of spliced mRNAs.
Figure 5: Timeline of morphological, cellular and molecular defects in embryos depleted of coilin.
Figure 6: Purified human snRNPs rescue the viability of coilin morphants.
Figure 7: Individual mature human snRNPs bypass the need for coilin, and snRNAs are not sufficient.

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  • 04 April 2010

    In the version of this article initially published online, the second sentence of the abstract should have read “Coilin is not a snRNP protein but concentrates snRNPs and their assembly intermediates in Cajal bodies (CBs).”. The error has been corrected for the print, PDF and HTML versions of this article.

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Acknowledgements

The authors thank D. Stanek, J. Geiger and C.-P. Heisenberg for their early involvement in this project and comments on the manuscript, K. Simons and M. Zerial for helpful discussions, G. Morris (Keele Univ.) for the gift of 1F1 antibody, I. Öchsner for her help with snRNA preparations, and the Max Planck Institute of Molecular Cell Biology fish facility. This work was supported by the Max Planck Society (K.M.N., A.C.O. and R.L.) and grants from the German Research Foundation (NE909/2-1 to K.M.N.) and the European Commission (EURASNET-518238 to K.M.N. and R.L.).

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Authors and Affiliations

  1. Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

    Magdalena Strzelecka, Andrew C Oates & Karla M Neugebauer

  2. Max Planck Institute of Biophysical Chemistry, Göttingen, Germany

    Simon Trowitzsch, Gert Weber & Reinhard Lührmann

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  1. Magdalena Strzelecka
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Contributions

M.S., R.L., A.C.O. and K.M.N. designed the experiments; S.T. and G.W. purified snRNPs and snRNAs; M.S. and K.M.N. performed the experiments; M.S. analyzed the data; M.S., A.C.O. and K.M.N. wrote the paper.

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Correspondence to Karla M Neugebauer.

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Strzelecka, M., Trowitzsch, S., Weber, G. et al. Coilin-dependent snRNP assembly is essential for zebrafish embryogenesis. Nat Struct Mol Biol 17, 403–409 (2010). https://doi.org/10.1038/nsmb.1783

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