Ribozyme-Catalyzed Transcription of an Active Ribozyme
Abstract
A critical event in the origin of life is thought to have been the emergence of an RNA molecule capable of replicating a primordial RNA “genome.” Here we describe the evolution and engineering of an RNA polymerase ribozyme capable of synthesizing RNAs of up to 95 nucleotides in length. To overcome its sequence dependence, we recombined traits evolved separately in different ribozyme lineages. This yielded a more general polymerase ribozyme that was able to synthesize a wider spectrum of RNA sequences, as we demonstrate by the accurate synthesis of an enzymatically active RNA, a hammerhead endonuclease ribozyme. This recapitulates a central aspect of an RNA-based genetic system: the RNA-catalyzed synthesis of an active ribozyme from an RNA template.
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References and Notes
1
R. Gesteland, T. R. Cech, J. F. Atkins, Eds., RNA World: The Nature of Modern RNA Suggests a Prebiotic RNA World (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, ed. 3, 2005).
2
Valadkhan S., Manley J. L., Splicing-related catalysis by protein-free snRNAs. Nature 413, 701 (2001).
3
Mandal M., Boese B., Barrick J. E., Winkler W. C., Breaker R. R., Riboswitches control fundamental biochemical pathways in Bacillus subtilis and other bacteria. Cell 113, 577 (2003).
4
Nissen P., Hansen J., Ban N., Moore P. B., Steitz T. A., The structural basis of ribosome activity in peptide bond synthesis. Science 289, 920 (2000).
5
Tuerk C., Gold L., Systematic evolution of ligands by exponential enrichment: RNA ligands to bacteriophage T4 DNA polymerase. Science 249, 505 (1990).
6
Ellington A. D., Szostak J. W., In vitro selection of RNA molecules that bind specific ligands. Nature 346, 818 (1990).
7
Johnston W. K., Unrau P. J., Lawrence M. S., Glasner M. E., Bartel D. P., RNA-catalyzed RNA polymerization: Accurate and general RNA-templated primer extension. Science 292, 1319 (2001).
8
Bartel D. P., Szostak J. W., Isolation of new ribozymes from a large pool of random sequences. Science 261, 1411 (1993).
9
Ekland E. H., Szostak J. W., Bartel D. P., Structurally complex and highly active RNA ligases derived from random RNA sequences. Science 269, 364 (1995).
10
Ekland E. H., Bartel D. P., RNA-catalysed RNA polymerization using nucleoside triphosphates. Nature 382, 373 (1996).
11
Materials and methods are available as supporting material on Science Online.
12
Joyce G. F., Directed evolution of nucleic acid enzymes. Annu. Rev. Biochem. 73, 791 (2004).
13
Wilson D. S., Szostak J. W., In vitro selection of functional nucleic acids. Annu. Rev. Biochem. 68, 611 (1999).
14
Tawfik D. S., Griffiths A. D., Man-made cell-like compartments for molecular evolution. Nat. Biotechnol. 16, 652 (1998).
15
Lawrence M. S., Bartel D. P., Processivity of ribozyme-catalyzed RNA polymerization. Biochemistry 42, 8748 (2003).
16
Zaher H. S., Unrau P. J., Selection of an improved RNA polymerase ribozyme with superior extension and fidelity. RNA 13, 1017 (2007).
17
Shechner D. M., et al., Crystal structure of the catalytic core of an RNA-polymerase ribozyme. Science 326, 1271 (2009).
18
Attwater J., Wochner A., Pinheiro V. B., Coulson A., Holliger P., Ice as a protocellular medium for RNA replication. Nat. Commun. 1, (2010).
19
Zhao H., Giver L., Shao Z., Affholter J. A., Arnold F. H., Molecular evolution by staggered extension process (StEP) in vitro recombination. Nat. Biotechnol. 16, 258 (1998).
20
McCall M. J., et al., Small, efficient hammerhead ribozymes. Mol. Biotechnol. 14, 5 (2000).
21
Wang Q. S., Cheng L. K., Unrau P. J., Characterization of the B6.61 polymerase ribozyme accessory domain. RNA 17, 469 (2011).
22
Shine J., Dalgarno L., Determinant of cistron specificity in bacterial ribosomes. Nature 254, 34 (1975).
23
Weiner A. M., Maizels N., tRNA-like structures tag the 3′ ends of genomic RNA molecules for replication: Implications for the origin of protein synthesis. Proc. Natl. Acad. Sci. U.S.A. 84, 7383 (1987).
24
Eigen M., Selforganization of matter and the evolution of biological macromolecules. Naturwissenschaften 58, 465 (1971).
25
Kun A., Santos M., Szathmáry E., Real ribozymes suggest a relaxed error threshold. Nat. Genet. 37, 1008 (2005).
26
Joyce G. F., A glimpse of biology’s first enzyme. Science 315, 1507 (2007).
27
Zuker M., Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 31, 3406 (2003).
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Published In
Science
Volume 332 | Issue 6026
8 April 2011
8 April 2011
Copyright
Copyright © 2011, American Association for the Advancement of Science.
Submission history
Received: 22 November 2010
Accepted: 15 February 2011
Published in print: 8 April 2011
Acknowledgments
We are grateful to M. Daly for assistance with FACS and to our colleagues V. B. Pinheiro, C. Cozens, A. Taylor, and J. Sutherland for helpful discussions. This work was supported by the Medical Research Council (MRC) program grant MC_US_A024_0014.
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