Abstract
Transcription factor IIIA (TFIIIA) is required for eukaryotic synthesis of 5S ribosomal RNA by RNA polymerase III. Here we report the discovery of a structured RNA element with clear resemblance to 5S rRNA that is conserved within TFIIIA precursor mRNAs from diverse plant lineages. TFIIIA protein expression is controlled by alternative splicing of the exon containing the plant 5S rRNA mimic (P5SM). P5SM triggers exon skipping upon binding of ribosomal protein L5, a natural partner of 5S rRNA, which demonstrates the functional adaptation of its structural mimicry. As the exon-skipped splice product encodes full-length TFIIIA protein, these results reveal a ribosomal protein–mRNA interaction that is involved in 5S rRNA synthesis and has implications for cross-coordination of ribosomal components. This study also provides insight into the origin and function of a newfound class of structured RNA that regulates alternative splicing.
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Acknowledgements
We thank K. Corbino, K. Leppek and A. Westermann for technical assistance, Z. Weinberg for advice on bioinformatics, E. Puerta-Fernandez for advice on gel-shift assays and A. Roth for critical reading of the manuscript. M.C.H. is supported by a Career Award at the Scientific Interface from the Burroughs Wellcome Fund, and A.W. was supported by the German Research Foundation while in the Breaker laboratory. The research was supported by a grant from the US National Institutes of Health (GM068819). RNA science in the Breaker laboratory also is supported by the Howard Hughes Medical Institute.
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M.C.H. conceived of and led project, performed bioinformatics analysis, designed and carried out in vitro structural probing and binding experiments, and wrote the manuscript. A.W. designed and carried out RT-PCR and qRT-PCR analyses and in vivo reporter assays, and edited the manuscript. R.R.B. advised on the project and edited the manuscript. All authors contributed to discussions regarding the data and their interpretation.
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Supplementary Figures 1–11, Supplementary Table 1, Supplementary Methods (PDF 8796 kb)
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Hammond, M., Wachter, A. & Breaker, R. A plant 5S ribosomal RNA mimic regulates alternative splicing of transcription factor IIIA pre-mRNAs. Nat Struct Mol Biol 16, 541–549 (2009). https://doi.org/10.1038/nsmb.1588
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DOI: https://doi.org/10.1038/nsmb.1588