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Structural and biochemical basis for misfolded RNA recognition by the Ro autoantigen

Nature Structural & Molecular Biology volume 13, pages 10021009 (2006) | Download Citation

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Abstract

The Ro autoantigen is ring-shaped, binds misfolded noncoding RNAs and is proposed to function in quality control. Here we determine how Ro interacts with misfolded RNAs. Binding of Ro to misfolded precursor (pre)-5S ribosomal RNA requires a single-stranded 3′ end and helical elements. As mutating most sequences of the helices and tail results in modest decreases in binding, Ro may be able to associate with a range of RNAs. Ro binds several other RNAs that contain single-stranded tails. A crystal structure of Ro bound to a misfolded pre-5S rRNA fragment reveals that the tail inserts into the cavity, while a helix binds on the surface. Most contacts of Ro with the helix are to the backbone. Mutagenesis reveals that Ro has an extensive RNA-binding surface. We propose that Ro uses this surface to scavenge RNAs that fail to bind their specific RNA-binding proteins.

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Acknowledgements

We thank A. Alexandrov, S. Sim, E. Ullu and E. Wurtmann for comments on the manuscript and S. Baserga (Yale University) and J. Steitz (Yale University) for gifts of plasmids. We are grateful to the staff at beamline X25 at the Brookhaven National Laboratory for assistance with data collection. A.J.S. was supported by a fellowship from the Arthritis Foundation. This work was funded by grants from the US National Institutes of Health (R01-GM073863 to S.L.W. and R01-GM70521 to K.M.R.) and the Pew Charitable Trust (to K.M.R.).

Author information

Author notes

    • Adam J Stein

    Present address: Department of Biochemistry and Molecular Biology, Rosalind Franklin University of Medicine and Science, North Chicago, Illinois 60064, USA.

Affiliations

  1. Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

    • Gabriele Fuchs
    • , Adam J Stein
    • , Chunmei Fu
    • , Karin M Reinisch
    •  & Sandra L Wolin
  2. Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510, USA.

    • Sandra L Wolin

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Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Karin M Reinisch or Sandra L Wolin.

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    Supplementary Fig. 1

    Difference electron density maps for RNA.

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DOI

https://doi.org/10.1038/nsmb1156