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Originally published In Press as doi:10.1074/jbc.M001460200 on May 31, 2000

J. Biol. Chem., Vol. 275, Issue 33, 25814-25819, August 18, 2000
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A 3'-5' Exonuclease in Human Leukemia Cells
IMPLICATIONS FOR RESISTANCE TO 1-beta -D-ARABINOFURANOSYLCYTOSINE AND 9-beta -D-ARABINOFURANOSYL-2-FLUOROADENINE 5'-MONOPHOSPHATE*

Violetta SkalskiDagger , Kevin R. Brown, Bo Yon Choi, Zhen-Yuan Lin, and Shali Chen

From the Division of Experimental Therapeutics, Ontario Cancer Institute, Princess Margaret Hospital and the Department of Medical Biophysics, University of Toronto, Toronto, Ontario M5G 2M9, Canada

A 3'-5' exonuclease that excises the nucleotide analogs 1-beta -D-arabinofuranosylcytosine monophosphate and 9-beta -D-arabinofuranosyl-2-fluoroadenine 5'-monophosphate incorporated at 3' ends of DNA was purified from the nuclei of: 1) primary human chronic lymphocytic leukemia cells, 2) primary and established human acute myeloblastic leukemia cells, and 3) lymphocytes obtained from healthy individuals. The activity of this nuclear exonuclease (exoN) is elevated approximately 6-fold in 1-beta -D-arabinofuranosylcytosine-resistant leukemia cells as compared with drug-sensitive cells, and it differs between two healthy individuals and among three leukemia patients. exoN is a 46-kDa monomer, requires 50 mM KCl and 1 mM magnesium for optimal activity, and shows a preference for single-stranded over duplex DNA. Its physical and enzymatic properties indicate that exoN is a previously uncharacterized enzyme whose activity may confer resistance to clinical nucleoside analogs in leukemia cells.

* This work was supported by Medical Research Council Grant MT-14350 (to V. S.) and an Operating Grant from the Leukemia Research Fund of Canada (to V. S.).The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed: Div. of Experimental Therapeutics, Ontario Cancer Inst., Princess Margaret Hospital, 620 University Ave., Toronto, Ontario M5G 2M9, Canada. Tel.: 416-946-2980; Fax: 416-946-2984; E-mail: skalski@oci.utoronto.ca.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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