MolPharm

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text (PDF)
Right arrow Submit a response
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Hentosh, P.
Right arrow Articles by Grippo, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Hentosh, P.
Right arrow Articles by Grippo, P.

Template 2-chloro-2'-deoxyadenosine monophosphate inhibits in vitro DNA synthesis

P Hentosh and P Grippo

Department of Pharmacology and Molecular Biology, Chicago Medical School, North Chicago, Illinois 60064.

2'-Chloro-2'-deoxyadenosine triphosphate (cladribine), a purine nucleotide analog and potent antileukemic agent, was enzymatically incorporated into 98-base oligomers in place of dATP to investigate the molecular consequences of 2-chloroadenine (CIAde) in DNA. We have used the resultant oligomers as templates for purified DNA polymerases, to compare the rate and extent of in vitro DNA synthesis; the sites of polymerase pausing, if any; and the effects of increasing deoxyribonucleoside triphosphate (dNTP) concentrations on synthetic reactions. Compared with control template, CIAde-containing DNA strikingly reduced the overall amount and rate of chain elongation by human polymerase beta and Klenow fragment. Distinct pause sites, which were polymerase dependent, occurred primarily one or two bases before or just after nucleotide incorporation opposite template CIAde. Human polymerase alpha and phage T4 DNA polymerase likewise exhibited reduced synthesis on CIAde-substituted templates. Bypassing of CIAde residues was possible only at higher dNTP concentrations, with approximately 20- and 50-fold greater dNTP concentrations being required for synthesis beyond CIAde sites, compared with adenine residues, by polymerase alpha and beta, respectively. These results suggest that CiAde residues located within cellular template DNA may inhibit daughter strand synthesis and thus contribute to the cytotoxic effects of the drug.

Volume 45, Issue 5, pp. 955-961, 05/01/1994
Copyright © 1994 by American Society for Pharmacology and Experimental Therapeutics




This article has been cited by other articles:


Home page
 Mol. Pharmacol.Home page
W. B. Parker, S. C. Shaddix, L. M. Rose, D. S. Shewach, L. W. Hertel, J. A. Secrist III, J. A. Montgomery, and L. L. Bennett Jr.
Comparison of the Mechanism of Cytotoxicity of 2-Chloro-9-(2-deoxy-2-fluoro-beta -D-arabinofuranosyl)adenine, 2-Chloro-9-(2-deoxy-2-fluoro-beta -D-ribofuranosyl)adenine, and 2-Chloro-9-(2-deoxy-2,2-difluoro-beta -D-ribofuranosyl)adenine in CEM Cells
Mol. Pharmacol., March 1, 1999; 55(3): 515 - 520.
[Abstract] [Full Text]

Home page
 Proc. Natl. Acad. Sci. USAHome page
L. M. Leoni, Q. Chao, H. B. Cottam, D. Genini, M. Rosenbach, C. J. Carrera, I. Budihardjo, X. Wang, and D. A. Carson
Induction of an apoptotic program in cell-free extracts by 2-chloro-2'-deoxyadenosine 5'-triphosphate and cytochrome c
PNAS, August 4, 1998; 95(16): 9567 - 9571.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
All ASPET Journals Molecular Pharmacology Pharmacological Reviews
Molecular Interventions Drug Metabolism and Disposition

Copyright © 1994 by the American Society for Pharmacology and Experimental Therapeutics