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  • Title: Structure-function relationship of hammerhead ribozymes as probed by 2'-modifications.
    Author: Pieken WA, Olsen DB, Aurup H, Williams DM, Heidenreich O, Benseler F, Eckstein F.
    Journal: Nucleic Acids Symp Ser; 1991; (24):51-3. PubMed ID: 1841379.
    Abstract:
    Hammerhead ribozymes containing 2'-fluoro- or 2'-aminonucleotides were prepared by automated chemical synthesis. Incorporation of 2'-fluorouridines, 2'-fluorocytidines or 2'-aminouridines did not appreciably decrease catalytic activity. The presence of 2'-aminocytidines, however, reduced the activity about 20-fold. No catalytic activity could be measured for ribozymes in which all adenosines were replaced by the 2'-fluoro analogue in presence of MgCl2. No single position could be found responsible for this loss of activity. In an attempt to construct ribozymes to hydrolyse HIV-RNA in the 5'-LTR region several constructs were tested on synthetic substrate as well as on run-off transcripts of about 1000 nucleotides length.
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