Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.

Science 20 December 1996:
Vol. 274. no. 5295, pp. 2065 - 2069
DOI: 10.1126/science.274.5295.2065
Prev | Table of Contents | Next

Reports

Capturing the Structure of a Catalytic RNA Intermediate: The Hammerhead Ribozyme

William G. Scott, * James B. Murray, John R. P. Arnold, Barry L. Stoddard, Aaron Klug

The crystal structure of an unmodified hammerhead RNA in the absence of divalent metal ions has been solved, and it was shown that this ribozyme can cleave itself in the crystal when divalent metal ions are added. This biologically active RNA fold is the same as that found previously for two modified hammerhead ribozymes. Addition of divalent cations at low pH makes it possible to capture the uncleaved RNA in metal-bound form. A conformational intermediate, having an additional Mg(II) bound to the cleavage-site phosphate, was captured by freeze-trapping the RNA at an active pH prior to cleavage. The most significant conformational changes were limited to the active site of the ribozyme, and the changed conformation requires only small additional movements to reach a proposed transition-state.

W. G. Scott and A. Klug, MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, England.
J. B. Murray and J.R.P. Arnold, Department of Biology, University of Leeds, Leeds LS2 9JT, England.
B. L. Stoddard, Fred Hutchinson Cancer Research Center, 1124 Columbia Street, Seattle, WA 98104, USA.
*   To whom correspondence should be addressed.

Read the Full Text


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Three-way RNA junctions with remote tertiary contacts: A recurrent and highly versatile fold.
M. d. l. Pena, D. Dufour, and J. Gallego (2009)
RNA 15, 1949-1964
   Abstract »    Full Text »    PDF »
Hammerhead redux: Does the new structure fit the old biochemical data?.
J. A. Nelson and O. C. Uhlenbeck (2008)
RNA 14, 605-615
   Abstract »    Full Text »    PDF »
Minimal and extended hammerheads utilize a similar dynamic reaction mechanism for catalysis.
J. A. Nelson and O. C. Uhlenbeck (2008)
RNA 14, 43-54
   Abstract »    Full Text »    PDF »
Effects of the trinucleotide preceding the self-cleavage site on eggplant latent viroid hammerheads: differences in co- and post-transcriptional self-cleavage may explain the lack of trinucleotide AUC in most natural hammerheads.
A. Carbonell, M. De la Pena, R. Flores, and S. Gago (2006)
Nucleic Acids Res. 34, 5613-5622
   Abstract »    Full Text »    PDF »
Binding of manganese(II) to a tertiary stabilized hammerhead ribozyme as studied by electron paramagnetic resonance spectroscopy.
N. KISSELEVA, A. KHVOROVA, E. WESTHOF, and O. SCHIEMANN (2005)
RNA 11, 1-6
   Abstract »    Full Text »    PDF »
Artificial tertiary motifs stabilize trans-cleaving hammerhead ribozymes under conditions of submillimolar divalent ions and high temperatures.
V. SAKSMERPROME, M. ROYCHOWDHURY-SAHA, S. JAYASENA, A. KHVOROVA, and D. H. BURKE (2004)
RNA 10, 1916-1924
   Abstract »    Full Text »    PDF »
Global structure of a DNA three-way junction by solution NMR: towards prediction of 3H fold.
B. Wu, F. Girard, B. van Buuren, J. Schleucher, M. Tessari, and S. Wijmenga (2004)
Nucleic Acids Res. 32, 3228-3239
   Abstract »    Full Text »    PDF »
Folding of the natural hammerhead ribozyme is enhanced by interaction of auxiliary elements.
J. C. PENEDO, T. J. WILSON, S. D. JAYASENA, A. KHVOROVA, and D. M.J. LILLEY (2004)
RNA 10, 880-888
   Abstract »    Full Text »    PDF »
Altered structural fluctuations in duplex RNA versus DNA: a conformational switch involving base pair opening.
Y. Pan and A. D. MacKerell Jr (2003)
Nucleic Acids Res. 31, 7131-7140
   Abstract »    Full Text »    PDF »
Structural Rearrangements of the 10-23 DNAzyme to {beta}3 Integrin Subunit mRNA Induced by Cations and Their Relations to the Catalytic Activity.
M. Cieslak, J. Szymanski, R. W. Adamiak, and C. S. Cierniewski (2003)
J. Biol. Chem. 278, 47987-47996
   Abstract »    Full Text »    PDF »
A crystallographic study of the binding of 13 metal ions to two related RNA duplexes.
E. Ennifar, P. Walter, and P. Dumas (2003)
Nucleic Acids Res. 31, 2671-2682
   Abstract »    Full Text »    PDF »
New Anti-Human Immunodeficiency Virus Type 1 6-Aminoquinolones: Mechanism of Action.
C. Parolin, B. Gatto, C. Del Vecchio, T. Pecere, E. Tramontano, V. Cecchetti, A. Fravolini, S. Masiero, M. Palumbo, and G. Palu (2003)
Antimicrob. Agents Chemother. 47, 889-896
   Abstract »    Full Text »    PDF »


To Advertise     Find Products

Science. ISSN 0036-8075 (print), 1095-9203 (online)