UC Santa Cruz
Capturing hammerhead ribozyme structures in action by modulating general base catalysis
- Author(s): Chi, YI
- Martick, M
- Lares, M
- Kim, R
- Scott, WG
- Kim, SH
- et al.
Published Web Locationhttps://doi.org/10.1371/journal.pbio.0060234
We have obtained precatalytic (enzyme-substrate complex) and postcatalytic (enzyme-product complex) crystal structures of an active full-length hammerhead RNA that cleaves in the crystal. Using the natural satellite tobacco ringspot virus hammerhead RNA sequence, the self-cleavage reaction was modulated by substituting the general base of the ribozyme, G12, with A12, a purine variant with a much lower pKathat does not significantly perturb the ribozyme's atomic structure. The active, but slowly cleaving, ribozyme thus permitted isolation of enzyme-substrate and enzyme-product complexes without modifying the nucleophile or leaving group of the cleavage reaction, nor any other aspect of the substrate. The predissociation enzyme-product complex structure reveals RNA and metal ion interactions potentially relevant to transition-state stabilization that are absent in precatalytic structures. © 2008 Chi et al.
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