Phosphoester-transfer mechanism of an RNA-cleaving acidic deoxyribozyme revealed by radioactivity tracking and enzymatic digestion.
A convenient method involving 32P-labeling of an RNA substrate at the cleavage site and subsequent enzymatic digestion of cleavage products via phosphatases reveals that pH4DZ1--an RNA-cleaving deoxyribozyme with optimal activity at pH 4--forms a 5'-cleavage fragment with 2',3'-cyclic phosphate group and a 3'-cleavage fragment with 5'-OH group.
Kandadai
Srinivas A
SA
Department of Biochemistry and Biomedical Sciences, Department of Chemistry, McMaster University, 1280 Main St. W. Hamilton, ON L8N 3Z5, Canada.
Chiuman
William
W
Li
Yingfu
Y
D021881:DNA, Catalytic / Q000737:chemistry*; D004789:Enzyme Activation; D006863:Hydrogen-Ion Concentration; D007553:Isotope Labeling; D010755:Organophosphates / Q000737:chemistry*; D010761:Phosphorus Radioisotopes / Q000737:chemistry; D012313:RNA / Q000737:chemistry*
D021881:DNA, Catalytic; D010755:Organophosphates; D010761:Phosphorus Radioisotopes; D012313:RNA