Study of a hammerhead ribozyme containing 2'-modified adenosine residues.
The improved synthesis of 2'-fluoro-2'-deoxyadenosine (2'-FA) starting from adenosine is described. This compound was converted to the phosphoramidite and incorporated into a hammerhead ribozyme RNA with the use of automated RNA synthesis techniques. Ribozymes containing 2'-deoxy-adenosine (2'-dA) were prepared in a similar manner. A kinetic rate comparison of the unmodified ribozyme with two ribozymes that had every adenosine replaced with 2'FA or 2'-dA revealed a large decrease in catalytic efficiency (kcat/Km) for the modified ribozymes resulting from a drop in kcat. The kinetic analysis of a number of partially substituted 2'-FA or 2'-dA containing hammerheads revealed that the decrease in activity was not associated with any particular residue but was the result of the accumulation of modified nucleosides within the structure.
Olsen
D B
DB
Max-Planck Institut für Experimentelle Medizin, Abteilung Chemie, Göttingen, FRG.
Benseler
F
F
Aurup
H
H
Pieken
W A
WA
Eckstein
F
F
D001483:Base Sequence; D003839:Deoxyadenosines / Q000737:chemistry* / Q000235:genetics; D007700:Kinetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D016337:RNA, Catalytic / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics; D013329:Structure-Activity Relationship
D003839:Deoxyadenosines; D016337:RNA, Catalytic; C030964:2'-fluoro-2'-deoxyadenosine