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 3â² 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.).
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Supplementary Fig. 1
Difference electron density maps for RNA. (PDF 1562 kb)
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Fuchs, G., Stein, A., Fu, C. et al. Structural and biochemical basis for misfolded RNA recognition by the Ro autoantigen. Nat Struct Mol Biol 13, 1002â1009 (2006). https://doi.org/10.1038/nsmb1156
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DOI: https://doi.org/10.1038/nsmb1156