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.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Autoantigens
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Base Sequence
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Binding Sites
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Crystallography, X-Ray
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Models, Molecular
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Molecular Sequence Data
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Mutagenesis
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Nuclease Protection Assays / methods
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Nucleic Acid Conformation*
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Oocytes / chemistry
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Oocytes / metabolism
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Protein Binding
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Protein Structure, Tertiary
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RNA 3' End Processing
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RNA Precursors / chemistry*
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RNA Precursors / genetics
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RNA Precursors / metabolism
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RNA, Ribosomal, 5S / chemistry*
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RNA, Ribosomal, 5S / genetics
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RNA, Ribosomal, 5S / metabolism
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RNA-Binding Proteins / chemistry
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RNA-Binding Proteins / genetics
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RNA-Binding Proteins / metabolism
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Ribonucleoproteins / chemistry*
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Ribonucleoproteins / genetics
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Ribonucleoproteins / metabolism*
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Xenopus laevis / embryology
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Xenopus laevis / genetics
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Xenopus laevis / metabolism
Substances
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Autoantigens
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RNA Precursors
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RNA, Ribosomal, 5S
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RNA-Binding Proteins
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Ribonucleoproteins
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SS-A antigen