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Nucleic Acids Research 2004 32(12):3531-3536; doi:10.1093/nar/gkh676
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Published online 6 July 2004

Nucleic Acids Research, Vol. 32 No. 12 © Oxford University Press 2004; all rights reserved

APE1 is the major 3'-phosphoglycolate activity in human cell extracts

Jason L. Parsons, Irina I. Dianova and Grigory L. Dianov*

MRC Radiation and Genome Stability Unit, Harwell, Oxfordshire OX11 0RD, UK

* To whom correspondence should be addressed. Tel: +44 1235 841 134; Fax: +44 1235 841 200; Email: g.dianov{at}har.mrc.ac.uk

Received April 30, 2004; Revised and Accepted June 9, 2004

DNA strand breaks containing 3'-phosphoglycolate (3'-PG) ends are the major lesions induced by ionizing radiation. The repair of this lesion is not completely understood and several activities are thought to be involved in processing of 3'-PG ends. In this study we examined activities in human whole cell extracts (WCE) responsible for removal of 3'-PG. Using a radiolabelled oligonucleotide containing a single nucleotide gap with internal 5'-phosphate and 3'-PG ends, we demonstrate that the major 3'-PG activity in human WCE is Mg2+ dependent and that this activity co-purifies with AP endonuclease 1 (APE1) over phosphocellulose and gel filtration chromatography. Furthermore, immunodepletion of APE1 from active gel filtration fractions using APE1 specific antibodies reveals that the major activity against 3'-PG in human WCE is APE1.


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