Structure-function relationship of hammerhead ribozymes as probed by 2'-modifications.
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.
Pieken
W A
WA
Max-Planck-Institut für Experimentelle Medizin, Göttingen, FRG.
Olsen
D B
DB
Aurup
H
H
Williams
D M
DM
Heidenreich
O
O
Benseler
F
F
Eckstein
F
F
D000998:Antiviral Agents / Q000138:chemical synthesis / Q000494:pharmacology; D001483:Base Sequence; D006678:HIV / Q000187:drug effects; D016325:HIV Long Terminal Repeat; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D009843:Oligoribonucleotides / Q000138:chemical synthesis* / Q000494:pharmacology; D016337:RNA, Catalytic / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D012367:RNA, Viral / Q000187:drug effects; D012265:Ribonucleotides
D000998:Antiviral Agents; D009843:Oligoribonucleotides; D016337:RNA, Catalytic; D012367:RNA, Viral; D012265:Ribonucleotides