title: Recognition of single-stranded DNA by the bacteriophage T4-induced type II topoisomerase.
abstract: The bacteriophage T4-induced type II DNA topoisomerase has been shown previously to make a reversible double strand break in DNA double helices. In addition, this enzyme is shown here to bind tightly and to cleave single-stranded DNA molecules. The evidence that the single-stranded DNA cleavage activity is intrinsic to the topoisomerase includes: 1) protein linkage to the 5' ends of the newly cleaved DNA; 2) coelution of essentially homogeneous topoisomerase and the DNA cleavage activity; 3) inhibition of both single-stranded DNA cleavage and double-stranded DNA relaxation by oxolinic acid; and 4) inhibition of duplex DNA relaxation by single-stranded DNA. The major cleavage sites on phi X174 viral DNA substrates have been mapped, and several cleavage sites analyzed to determine the exact nucleotide position of cleavage. Major cleavage sites are found very near the base of predicted hairpin helices in the single-stranded DNA substrates, suggesting that DNA secondary structure recognition is important in the cleavage reaction. On the other hand, there are also many weaker cleavage sites with no obvious sequence requirements. Many of the properties of the single-stranded DNA cleavage reaction examined here differ from those of the oxolinic acid-dependent, double-stranded DNA cleavage reaction catalyzed by the same enzyme.
mesh_terms: D000595:Amino Acid Sequence; D010584:Bacteriophage phi X 174; D001483:Base Sequence; D004262:DNA Restriction Enzymes; D004250:DNA Topoisomerases, Type II / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology; D008970:Molecular Weight; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D013379:Substrate Specificity; D013604:T-Phages / Q000201:enzymology*
chemical_list: D004277:DNA, Single-Stranded; D004262:DNA Restriction Enzymes; D004250:DNA Topoisomerases, Type II