Deletion analysis in the catalytic region of the 10-23 DNA enzyme.
In this study, the functional relevance of the core nucleotides of the RNA cleaving 10-23 DNA enzyme (DNAzyme) was investigated. Systematic deletion studies revealed that DNAzymes lacking thymine at position 8 (T8) retain catalytic activity comparable to that of the wild-type enzyme. Deletion of the adjacent cytosine at position 7 (C7) also resulted in a highly active enzyme and even the double deletion mutant C7/T8 displayed cleavage activity, although the catalytic rate under multiple turnover conditions was found to be reduced by one order of magnitude. The identification of non-essential nucleotides in the catalytic core might help to stabilize the DNAzyme against nucleolytic degradation and to overcome problems in elucidating its three-dimensional structure.
Zaborowska
Zaneta
Z
Institute of Biochemistry, Free University Berlin, Thielallee 63, 14195 Berlin, Germany.
Schubert
Steffen
S
Kurreck
Jens
J
Erdmann
Volker A
VA
D001483:Base Sequence / Q000235:genetics; D020134:Catalytic Domain / Q000235:genetics; D004252:DNA Mutational Analysis; D021881:DNA, Catalytic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D007477:Ions / Q000378:metabolism; D007700:Kinetics; D009690:Nucleic Acid Conformation; D012313:RNA / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics; D044366:Transition Temperature
D021881:DNA, Catalytic; D004277:DNA, Single-Stranded; D007477:Ions; C402582:RNA-cleaving DNA 10-23; D012313:RNA