Enzymic in vitro repair and chemical nature of DNA chain breaks induced by incorporated phosphorus-<sup>32</sup>p decay

Fodor, I; Matvienko, N.I.; Злотников, К. М.; Tanyashin, V. I.; Solonin, Alexander S.; Bayev, A.A.

doi:10.1093/nar/2.5.635

Cited by 4 publications

(1 citation statement)

References 16 publications

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“…The failure of the previous study to detect exonucleolytic cleavage of a 3′phosphate-terminated oligodeoxynucleotide by polymerase I (17) was probably due to the short size (a 7-mer) and low concentration of the substrate used and the inefficient nature of the reaction as well as the possible requirement for the presence of a dNTP. The observation that polymerase I and DNA ligase can repair 50% of X-ray-induced strand breaks (25) and 20-30% of 32 P-induced strand breaks (26) suggests that though polymerase I is able to process the 3′-phosphateterminated gaps, it is not able to process those terminating in a 3′-phosphoglycolate or other types of ends as well.…”

Section: Resultsmentioning

confidence: 99%

Phototriggered Formation and Repair of DNA Containing a Site-Specific Single Strand Break of the Type Produced by Ionizing Radiation or AP Lyase Activity

Zhang

Taylor

2000

Biochemistry

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DNA strand breaks are produced by a variety of agents and processes such as ionizing radiation, xenobiotics, oxidative metabolism, and enzymatic processing of DNA base damage. One of the major types of strand breaks produced by these processes is a single nucleotide gap terminating in 5'- and 3'-phosphates. Previously, we had developed a method for sequence-specifically producing such phosphate-terminated strand breaks in an oligodeoxynucleotide by way of two photochemically activated (caged) building blocks placed in tandem. We now report the design and synthesis of a single caged building block consisting of 1,3-(2-nitrophenyl)-1,3-propanediol, for producing phosphate-terminated strand breaks, and its use producing such a break at a specific site in a double-stranded circular DNA vector. To produce the site-specific break in a duplex vector, a primer containing the caged single strand break was extended opposite the single strand form of a circular DNA vector followed by enzymatic ligation and purification. The single strand break could then be formed in quantitative yield by irradiation of the vector with 365 nm light. In contrast to a previous study, it was found that the strand break can be repaired by Escherichia coli DNA polymerase I and E. coli DNA ligase alone, though less efficiently than in the presence of the 3'-phosphate processing enzyme E. coli endonuclease IV. Repair in the absence of endonuclease IV could be attributed to hydrolysis of the 3'-phosphate in the presence of dNTP and to a lesser extent to exonucleolytic removal of the 3'-phosphate-bearing terminal nucleotide by way of the 3' --> 5' exonuclease activity of polymerase I. This work demonstrates that specialized 3'-end processing enzymes such as endonuclease IV or exonuclease III are not absolutely required for repair of phosphate-terminated gaps. In addition to preparing single strand breaks, the caged building block described should also be useful for preparing double strand breaks and multiply damaged sites that might otherwise be difficult to prepare by other methods due to their lability.

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Section: Resultsmentioning

confidence: 99%

Phototriggered Formation and Repair of DNA Containing a Site-Specific Single Strand Break of the Type Produced by Ionizing Radiation or AP Lyase Activity

Zhang

Taylor

2000

Biochemistry

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Polynucleotide Kinase

Kleppe¹

1979

Advances in Enzymology - And Related Areas of Molecular Biology

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In vitro repair of radiation-induced strand breaks in DNA

Mitzel-Landbeck

Schütz

Hagen

1976

Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein

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Enzymic in vitro repair and chemical nature of DNA chain breaks induced by incorporated phosphorus-³²p decay

Cited by 4 publications

References 16 publications

Phototriggered Formation and Repair of DNA Containing a Site-Specific Single Strand Break of the Type Produced by Ionizing Radiation or AP Lyase Activity

Phototriggered Formation and Repair of DNA Containing a Site-Specific Single Strand Break of the Type Produced by Ionizing Radiation or AP Lyase Activity

Polynucleotide Kinase

In vitro repair of radiation-induced strand breaks in DNA

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Enzymic in vitro repair and chemical nature of DNA chain breaks induced by incorporated phosphorus-32p decay

Cited by 4 publications

References 16 publications

Phototriggered Formation and Repair of DNA Containing a Site-Specific Single Strand Break of the Type Produced by Ionizing Radiation or AP Lyase Activity

Phototriggered Formation and Repair of DNA Containing a Site-Specific Single Strand Break of the Type Produced by Ionizing Radiation or AP Lyase Activity

Polynucleotide Kinase

In vitro repair of radiation-induced strand breaks in DNA

Contact Info

Product

Resources

About

Enzymic in vitro repair and chemical nature of DNA chain breaks induced by incorporated phosphorus-³²p decay