Abstract
We report the structural basis for the modulation of an ATP-sensitive ribozyme that was engineered by modular rational design. This allosteric ribozyme is composed of two independently functioning domains, one a receptor for ATP and the other a self-cleaving ribozyme. When fused in the appropriate fashion, the conjoined aptamer-ribozyme construct functions as an allosteric ribozyme that is inhibited in the presence of ATP. The aptamer domain remains conformationally heterogeneous in the absence of ATP, but folds into a distinct structure upon ligand binding. This ATP-induced conformational change causes a reduction in catalytic activity of the adjacent ribozyme domain due to steric interference between the aptamer and ribozyme tertiary structures. This mechanism for structural and functional modulation of nucleic acids is one of several possible mechanisms by which the function of ribozymes could be specifically controlled by small effector molecules.
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- Arnott S., Hukins D. W., Dover S. D. Optimised parameters for RNA double-helices. Biochem Biophys Res Commun. 1972 Sep 26;48(6):1392–1399. doi: 10.1016/0006-291x(72)90867-4. [DOI] [PubMed] [Google Scholar]
- Bassi G. S., Møllegaard N. E., Murchie A. I., von Kitzing E., Lilley D. M. Ionic interactions and the global conformations of the hammerhead ribozyme. Nat Struct Biol. 1995 Jan;2(1):45–55. doi: 10.1038/nsb0195-45. [DOI] [PubMed] [Google Scholar]
- Breaker R. R. DNA aptamers and DNA enzymes. Curr Opin Chem Biol. 1997 Jun;1(1):26–31. doi: 10.1016/s1367-5931(97)80105-6. [DOI] [PubMed] [Google Scholar]
- Breaker R. R. DNA enzymes. Nat Biotechnol. 1997 May;15(5):427–431. doi: 10.1038/nbt0597-427. [DOI] [PubMed] [Google Scholar]
- Breaker Ronald R. In Vitro Selection of Catalytic Polynucleotides. Chem Rev. 1997 Apr 1;97(2):371–390. doi: 10.1021/cr960008k. [DOI] [PubMed] [Google Scholar]
- Carey J. Gel retardation at low pH resolves trp repressor-DNA complexes for quantitative study. Proc Natl Acad Sci U S A. 1988 Feb;85(4):975–979. doi: 10.1073/pnas.85.4.975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clouet-d'Orval B., Stage T. K., Uhlenbeck O. C. Neomycin inhibition of the hammerhead ribozyme involves ionic interactions. Biochemistry. 1995 Sep 5;34(35):11186–11190. doi: 10.1021/bi00035a025. [DOI] [PubMed] [Google Scholar]
- Dieckmann T., Suzuki E., Nakamura G. K., Feigon J. Solution structure of an ATP-binding RNA aptamer reveals a novel fold. RNA. 1996 Jul;2(7):628–640. [PMC free article] [PubMed] [Google Scholar]
- Draper D. E. Strategies for RNA folding. Trends Biochem Sci. 1996 Apr;21(4):145–149. [PubMed] [Google Scholar]
- Feig A. L., Scott W. G., Uhlenbeck O. C. Inhibition of the hammerhead ribozyme cleavage reaction by site-specific binding of Tb. Science. 1998 Jan 2;279(5347):81–84. doi: 10.1126/science.279.5347.81. [DOI] [PubMed] [Google Scholar]
- Forster A. C., Symons R. H. Self-cleavage of plus and minus RNAs of a virusoid and a structural model for the active sites. Cell. 1987 Apr 24;49(2):211–220. doi: 10.1016/0092-8674(87)90562-9. [DOI] [PubMed] [Google Scholar]
- Jiang F., Kumar R. A., Jones R. A., Patel D. J. Structural basis of RNA folding and recognition in an AMP-RNA aptamer complex. Nature. 1996 Jul 11;382(6587):183–186. doi: 10.1038/382183a0. [DOI] [PubMed] [Google Scholar]
- Koshland D. E., Jr, Némethy G., Filmer D. Comparison of experimental binding data and theoretical models in proteins containing subunits. Biochemistry. 1966 Jan;5(1):365–385. doi: 10.1021/bi00865a047. [DOI] [PubMed] [Google Scholar]
- Long D. M., Uhlenbeck O. C. Kinetic characterization of intramolecular and intermolecular hammerhead RNAs with stem II deletions. Proc Natl Acad Sci U S A. 1994 Jul 19;91(15):6977–6981. doi: 10.1073/pnas.91.15.6977. [DOI] [PMC free article] [PubMed] [Google Scholar]
- MONOD J., CHANGEUX J. P., JACOB F. Allosteric proteins and cellular control systems. J Mol Biol. 1963 Apr;6:306–329. doi: 10.1016/s0022-2836(63)80091-1. [DOI] [PubMed] [Google Scholar]
- MONOD J., JACOB F. Teleonomic mechanisms in cellular metabolism, growth, and differentiation. Cold Spring Harb Symp Quant Biol. 1961;26:389–401. doi: 10.1101/sqb.1961.026.01.048. [DOI] [PubMed] [Google Scholar]
- Miller W. A., Silver S. L. Alternative tertiary structure attenuates self-cleavage of the ribozyme in the satellite RNA of barley yellow dwarf virus. Nucleic Acids Res. 1991 Oct 11;19(19):5313–5320. doi: 10.1093/nar/19.19.5313. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Olive J. E., De Abreu D. M., Rastogi T., Andersen A. A., Mittermaier A. K., Beattie T. L., Collins R. A. Enhancement of Neurospora VS ribozyme cleavage by tuberactinomycin antibiotics. EMBO J. 1995 Jul 3;14(13):3247–3251. doi: 10.1002/j.1460-2075.1995.tb07327.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Osborne Scott E., Ellington Andrew D. Nucleic Acid Selection and the Challenge of Combinatorial Chemistry. Chem Rev. 1997 Apr 1;97(2):349–370. doi: 10.1021/cr960009c. [DOI] [PubMed] [Google Scholar]
- Peracchi A., Beigelman L., Usman N., Herschlag D. Rescue of abasic hammerhead ribozymes by exogenous addition of specific bases. Proc Natl Acad Sci U S A. 1996 Oct 15;93(21):11522–11527. doi: 10.1073/pnas.93.21.11522. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pley H. W., Flaherty K. M., McKay D. B. Three-dimensional structure of a hammerhead ribozyme. Nature. 1994 Nov 3;372(6501):68–74. doi: 10.1038/372068a0. [DOI] [PubMed] [Google Scholar]
- Porta H., Lizardi P. M. An allosteric hammerhead ribozyme. Biotechnology (N Y) 1995 Feb;13(2):161–164. doi: 10.1038/nbt0295-161. [DOI] [PubMed] [Google Scholar]
- Pyle A. M., Green J. B. RNA folding. Curr Opin Struct Biol. 1995 Jun;5(3):303–310. doi: 10.1016/0959-440x(95)80091-3. [DOI] [PubMed] [Google Scholar]
- Sassanfar M., Szostak J. W. An RNA motif that binds ATP. Nature. 1993 Aug 5;364(6437):550–553. doi: 10.1038/364550a0. [DOI] [PubMed] [Google Scholar]
- Scott W. G., Finch J. T., Klug A. The crystal structure of an all-RNA hammerhead ribozyme: a proposed mechanism for RNA catalytic cleavage. Cell. 1995 Jun 30;81(7):991–1002. doi: 10.1016/s0092-8674(05)80004-2. [DOI] [PubMed] [Google Scholar]
- Scott W. G., Murray J. B., Arnold J. R., Stoddard B. L., Klug A. Capturing the structure of a catalytic RNA intermediate: the hammerhead ribozyme. Science. 1996 Dec 20;274(5295):2065–2069. doi: 10.1126/science.274.5295.2065. [DOI] [PubMed] [Google Scholar]
- Sigurdsson S. T., Tuschl T., Eckstein F. Probing RNA tertiary structure: interhelical crosslinking of the hammerhead ribozyme. RNA. 1995 Aug;1(6):575–583. [PMC free article] [PubMed] [Google Scholar]
- Tang J., Breaker R. R. Examination of the catalytic fitness of the hammerhead ribozyme by in vitro selection. RNA. 1997 Aug;3(8):914–925. [PMC free article] [PubMed] [Google Scholar]
- Tang J., Breaker R. R. Rational design of allosteric ribozymes. Chem Biol. 1997 Jun;4(6):453–459. doi: 10.1016/s1074-5521(97)90197-6. [DOI] [PubMed] [Google Scholar]
- Tuschl T., Eckstein F. Hammerhead ribozymes: importance of stem-loop II for activity. Proc Natl Acad Sci U S A. 1993 Aug 1;90(15):6991–6994. doi: 10.1073/pnas.90.15.6991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tuschl T., Gohlke C., Jovin T. M., Westhof E., Eckstein F. A three-dimensional model for the hammerhead ribozyme based on fluorescence measurements. Science. 1994 Nov 4;266(5186):785–789. doi: 10.1126/science.7973630. [DOI] [PubMed] [Google Scholar]
- Wank H., Rogers J., Davies J., Schroeder R. Peptide antibiotics of the tuberactinomycin family as inhibitors of group I intron RNA splicing. J Mol Biol. 1994 Mar 4;236(4):1001–1010. doi: 10.1016/0022-2836(94)90007-8. [DOI] [PubMed] [Google Scholar]
- von Ahsen U., Davies J., Schroeder R. Antibiotic inhibition of group I ribozyme function. Nature. 1991 Sep 26;353(6342):368–370. doi: 10.1038/353368a0. [DOI] [PubMed] [Google Scholar]
