Quantitative Assays for Uracil-DNA Glycosylase of High Sensitivity

Overview

Journal Biochem Cell Biol

Specialties Biochemistry
Cell Biology

Date 1988 Feb 1

PMID 3370147

Citations 4

Authors

A R Morgan

J Chlebek

Affiliations

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Abstract

We have developed a sensitive fluorometric assay using bisulfite deaminated (C----U), covalently-closed circular PM2 DNA as the substrate. We describe a reliable way to prepare this substrate without nicking the PM2 DNA. Methods, which depend on toluenization of the cells, are described for reproducibly and quantitatively assaying uracil-DNA glycosylase. The sensitivity is such that only 200 EL4 mouse thymoma cells or 30,000 Escherichia coli cells are needed for each point in a kinetic assay.

Citing Articles

Vaccinia virus uracil DNA glycosylase has an essential role in DNA synthesis that is independent of its glycosylase activity: catalytic site mutations reduce virulence but not virus replication in cultured cells.

De Silva F, Moss B J Virol. 2002; 77(1):159-66.

PMID: 12477821 PMC: 140647. DOI: 10.1128/jvi.77.1.159-166.2003.

Characterization of uracil-DNA glycosylase activity from Trypanosoma cruzi and its stimulation by AP endonuclease.

Gallego C, Ruiz-Perez L, Gonzalez-Pacanowska D Nucleic Acids Res. 2001; 29(7):1549-55.

PMID: 11266557 PMC: 31275. DOI: 10.1093/nar/29.7.1549.

Mutations in active-site residues of the uracil-DNA glycosylase encoded by vaccinia virus are incompatible with virus viability.

Ellison K, Peng W, McFadden G J Virol. 1996; 70(11):7965-73.

PMID: 8892920 PMC: 190869. DOI: 10.1128/JVI.70.11.7965-7973.1996.

Identification of a poxvirus gene encoding a uracil DNA glycosylase.

Upton C, Stuart D, McFadden G Proc Natl Acad Sci U S A. 1993; 90(10):4518-22.

PMID: 8389453 PMC: 46543. DOI: 10.1073/pnas.90.10.4518.