A unique endonuclease from Crithidia fasciculata which recognizes a bend in the DNA helix. Specificity of the cleavage reaction.
The introduction of a single nick in DNA circles by Crithidia fasciculata nicking enzyme (Shlomai, J., and Linial, M. (1986) J. Biol. Chem. 261, 16219-16225) requires the presence of a bent structure in the DNA helix. However, the sequence directing the local bending of the DNA helix is not per se a preferred site for nicking by the enzyme. No extensive sequence specificity is involved in defining the cleavage site for C. fasciculata nicking enzyme in the duplex circular DNA substrate. However, the abundance of A and T residues is significantly high at both the 3' and the 5' termini generated at the nicked site. Nicking of the sequence-directed bent fragment from C. fasciculata kinetoplast DNA minicircles correlates with the periodicity determined by the unique nucleotide distribution in the bent sequence, reflected in its thermodynamic parameters. Occurrence of nicking is best correlated with the predicted minima of the melting temperature and delta G profiles, as well as with A and T dinucleotide sequences at the nicked site, in both the supercoiled and the relaxed sequence-directed bent DNA substrates. The potential role of the bend-dependent nicking reaction in the replication of kinetoplast DNA minicircles is discussed.
Linial
M
M
Kuvin Centre for the Study of Infectious and Tropical Diseases, Hebrew University, Hadassah Medical School, Jerusalem, Israel.
Shlomai
J
J
D000818:Animals; D003421:Crithidia / Q000201:enzymology*; D004247:DNA; D004262:DNA Restriction Enzymes / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D007700:Kinetics; D009690:Nucleic Acid Conformation; D010957:Plasmids; D013379:Substrate Specificity
D004247:DNA; D004262:DNA Restriction Enzymes; D003850:Deoxyribonuclease I