Abstract
The covalent joining of topoisomerases to DNA is normally a transient step in the reaction cycle of these important enzymes. However, under a variety of circumstances, the covalent complex is converted to a long-lived or dead-end product that can result in chromosome breakage and cell death. We have discovered and partially purified an enzyme that specifically cleaves the chemical bond that joins the active site tyrosine of topoisomerases to the 3' end of DNA. The reaction products made by the purified enzyme on a variety of model substrates indicate that the enzyme cleanly hydrolyzes the tyrosine-DNA phosphodiester linkage, thereby liberating a DNA terminated with a 3' phosphate. The wide distribution of this phosphodiesterase in eukaryotes and its specificity for tyrosine linked to the 3' end but not the 5' end of DNA suggest that it plays a role in the repair of DNA trapped in complexes involving eukaryotic topoisomerase I.
MeSH Terms
Binding Sites
DNA/chemistry,metabolism
DNA Repair
DNA Topoisomerases, Type I/chemistry,metabolism
Kinetics
Models, Chemical
Molecular Weight
Oligodeoxyribonucleotides/chemical synthesis,chemistry,metabolism
Phosphoric Diester Hydrolases/isolation & purification,metabolism
Saccharomyces cerevisiae/enzymology
Substrate Specificity
Tyrosine
Chemicals
Oligodeoxyribonucleotides
Tyrosine
DNA
Phosphoric Diester Hydrolases
DNA Topoisomerases, Type I
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Yang S W
Laboratory of Molecular Biology, National Institute of Mental Health, Bethesda, MD 20892-4034, USA.
Burgin A B
Huizenga B N
Robertson C A
Yao K C
Nash H A
References (25)
25 references, click to expand
-
A novel suicide substrate for DNA topoisomerases and site-specific recombinases.
Nucleic Acids Res. 1995 Aug 11;23(15):2973-9
PMID: 7659520
-
5'phosphorylation of DNA in mammalian cells: identification of a polymin P-precipitable polynucleotide kinase.
J Cell Biochem. 1995 May;58(1):115-31
PMID: 7642718
-
Human DNA topoisomerase I-mediated cleavages stimulated by ultraviolet light-induced DNA damage.
J Biol Chem. 1996 Mar 22;271(12):6978-86
PMID: 8636127
-
DNA topoisomerases.
Annu Rev Biochem. 1996;65:635-92
PMID: 8811192
-
Preclinical and clinical development of camptothecins.
Adv Pharmacol. 1994;29B:51-72
PMID: 8996601
-
Mechanisms of topoisomerase I inhibition by anticancer drugs.
Adv Pharmacol. 1994;29B:73-92
PMID: 8996602
-
Viral integration and excision: structure of the lambda att sites.
Science. 1977 Sep 16;197(4309):1147-60
PMID: 331474
-
DNA structural features that lead to strand breakage by eukaryotic type-I topoisomerase.
Cold Spring Harb Symp Quant Biol. 1984;49:435-42
PMID: 6099251
-
Chromatin 3'-phosphatase/5'-OH kinase cannot transfer phosphate from 3' to 5' across a strand nick in DNA.
Nucleic Acids Res. 1986 Oct 24;14(20):8103-10
PMID: 3022244
-
Site-specific recombination intermediates trapped with suicide substrates.
Cell. 1987 Aug 28;50(5):779-88
PMID: 3040260
-
DNA topoisomerase-targeting antitumor drugs can be studied in yeast.
Proc Natl Acad Sci U S A. 1988 Oct;85(20):7501-5
PMID: 2845409
-
Minimal DNA duplex requirements for topoisomerase I-mediated cleavage in vitro.
J Biol Chem. 1990 Jul 25;265(21):12529-35
PMID: 2165067
-
Isolation of yeast nuclei.
Methods Enzymol. 1991;194:735-49
PMID: 2005821
-
Symmetry in the mechanism of bacteriophage lambda integrative recombination.
Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9642-6
PMID: 1409677
-
Ligation of synthetic activated DNA substrates by site-specific recombinases and topoisomerase I.
J Biol Chem. 1993 Feb 15;268(5):3683-9
PMID: 8381436
-
The covalent eukaryotic topoisomerase I-DNA intermediate catalyzes pH-dependent hydrolysis and alcoholysis.
J Biol Chem. 1994 Apr 15;269(15):11367-73
PMID: 8157668
-
Mammalian topoisomerase I has base mismatch nicking activity.
J Biol Chem. 1994 Jun 3;269(22):15498-504
PMID: 8195193
-
DNA topoisomerases: essential enzymes and lethal targets.
Annu Rev Pharmacol Toxicol. 1994;34:191-218
PMID: 8042851
-
Specific cleavage of model recombination and repair intermediates by the yeast Rad1-Rad10 DNA endonuclease.
Science. 1994 Sep 30;265(5181):2082-5
PMID: 8091230
-
Cellular determinants of sensitivity and resistance to DNA topoisomerase inhibitors.
Cancer Invest. 1994;12(5):530-42
PMID: 7922710
-
Architectural elements in nucleoprotein complexes: interchangeability of specific and non-specific DNA binding proteins.
EMBO J. 1994 Oct 3;13(19):4536-48
PMID: 7925295
-
Cellular regulation of mammalian DNA topoisomerases.
Adv Pharmacol. 1994;29A:167-89
PMID: 7826857
-
Topoisomerase I-related parameters and camptothecin activity in the colon carcinoma cell lines from the National Cancer Institute anticancer screen.
Cancer Res. 1995 May 15;55(10):2116-21
PMID: 7743511
-
SCT1 mutants suppress the camptothecin sensitivity of yeast cells expressing wild-type DNA topoisomerase I.
Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6299-303
PMID: 7603986
-
Topoisomerase poisons: harnessing the dark side of enzyme mechanism.
J Biol Chem. 1995 Sep 15;270(37):21429-32
PMID: 7665550