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Journal Article

Structural principles of RNA catalysis in a 2 '-5 ' lariat-forming ribozyme.

MPS-Authors
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Fohrer,  J.
Department of NMR Based Structural Biology, MPI for biophysical chemistry, Max Planck Society;

Fulltext (public)

1744103.pdf
(Publisher version), 4MB

Supplementary Material (public)

1744103_Supplement_1.pdf
(Supplementary material), 915KB

1744103_Supplement_2.pdb
(Supplementary material), 145KB

Citation

Carlomagno, T., Amata, I., Codutti, L., Falb, M., Fohrer, J., Masiewicz, P., et al. (2013). Structural principles of RNA catalysis in a 2 '-5 ' lariat-forming ribozyme. Journal of the American Chemical Society, 135(11), 4403-4411. doi:10.1021/ja311868t.


Cite as: http://hdl.handle.net/11858/00-001M-0000-000E-FC6C-A
Abstract
RNA-catalyzed lariat formation is present in both eukaryotes and prokaryotes. To date we lack structural insights into the catalytic mechanism of lariat-forming ribozymes. Here, we study an artificial 2'-5' AG1 lariat-forming ribozyme that shares the sequence specificity of lariat formation with the pre-mRNA splicing reaction. Using NMR, we solve the structure of the inactive state of the ribozyme in the absence of magnesium. The reaction center S'-guanosine appears to be part of a helix with an exceptionally widened major groove, while the lariat-forming A48 is looped out at the apex of a pseudoknot. The model of the active state built by mutational analysis, molecular modeling, and small-angle X-ray scattering suggests that A48 is recognized by a conserved adenosine, juxtaposed to the 5'-guanosine in one base-pair step distance, while the G1-N7 coordinates a magnesium ion essential for the activation of the nucleophile. Our findings offer implications for lariat formation in RNA enzymes including the mechanism of the recognition of the branch-site adenosine.