In vitro Selection of Self-Cleaving RNAs with a Low pH Optimum

RNAs that undergo a rapid site-specific cleavage at low pH have been selected by in vitro selection (the SELEX process). The cleavage does not require the addition of any divalent metal ions, and is in fact inhibited by divalent metal ions, spermine, or high concentrations of monovalent metal ions....

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1997-09, Vol.94 (20), p.10612-10617
Main Authors: Jayasena, Vineetha K., Gold, Larry
Format: Article
Language:English
Subjects:
Acetates
Biochemistry
Biological Sciences
Catalysis
Enzymes
Gels
Hydrogen-Ion Concentration
Hydrolysis
Metal ions
Metals - metabolism
Molecules
Nucleotides
Ribonucleic acid
RNA
RNA, Catalytic - metabolism
Room temperature
Sodium
Space life sciences
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Summary:RNAs that undergo a rapid site-specific cleavage at low pH have been selected by in vitro selection (the SELEX process). The cleavage does not require the addition of any divalent metal ions, and is in fact inhibited by divalent metal ions, spermine, or high concentrations of monovalent metal ions. This low pH catalyzed cleavage results in a 2′,3′-cyclic phosphate at the 3′end and a free hydroxyl at the 5′end. The reaction proceeds with a calculated rate of 1.1 min-1at room temperature in cacodylate buffer at pH 5.0. The rate of cleavage is dependent on the pH and shows an optimum around pH 4.0. The rate constant is independent of RNA concentration, indicating to an intramolecular reaction. Autocatalytic cleavage at low pH, in the absence of a metal ion requirement, adds to the reaction possibilities that may have existed on the prebiotic earth.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.94.20.10612