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Inhibition of Klenow fragment DNA polymerase on double-helical templates by oligonucleotide-directed triple-helix formation

Hacia, Joseph G. and Dervan, Peter B. and Wold, Barbara J. (1994) Inhibition of Klenow fragment DNA polymerase on double-helical templates by oligonucleotide-directed triple-helix formation. Biochemistry, 33 (20). pp. 6192-6200. ISSN 0006-2960. https://resolver.caltech.edu/CaltechAUTHORS:20160510-133746042

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Abstract

We have examined the capacity of oligonucleotide-directed triple helices to block the progress of primer extension by DNA polymerase. Occupancy of the major groove of a double-helical DNA substrate obstructed Klenow fragment progress at sites that map near the proximal boundary between duplex and triplex. Among a family of related third-strand oligonucleotides that all stably occupied the target duplex in the absence of polymerase, those forming longer triplexes were more effective polymerase inhibitors than shorter complexes. Kinetic analysis revealed that the triple-helical complex provided an effective blockade for times of at least 20 min. These observations provide the basis for considering and further dissecting repair DNA polymerase function and mechanism by using various defined local three-stranded DNA structures as probes.


Item Type:Article
Related URLs:
URLURL TypeDescription
http://dx.doi.org/10.1021/bi00186a019DOIArticle
http://pubs.acs.org/doi/abs/10.1021/bi00186a019PublisherArticle
ORCID:
AuthorORCID
Dervan, Peter B.0000-0001-8852-7306
Wold, Barbara J.0000-0003-3235-8130
Additional Information:© 1994 American Chemical Society. Received January 7, 1994; Revised Manuscript Received March 10, 1994. Abstract published in Advance ACS Abstracts, April 15, 1994. Supported by a predoctoral fellowship from the Howard Hughes Medical Institute to J.G.H. and by research grants from the NIH (GM35924) to P.B.D. and from the NIH (AR40780) to B.J.W. We gratefully acknowledge J. Campbell for valuable discussion and critical reading of the manuscript.
Funders:
Funding AgencyGrant Number
Howard Hughes Medical Institute (HHMI)UNSPECIFIED
NIHGM-35924
NIHAR40780
Issue or Number:20
Record Number:CaltechAUTHORS:20160510-133746042
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20160510-133746042
Official Citation:Inhibition of Klenow Fragment DNA Polymerase on Double-Helical Templates by Oligonucleotide-Directed Triple-Helix Formation Joseph G. Hacia, Peter B. Dervan, and Barbara J. Wold Biochemistry 1994 33 (20), 6192-6200 DOI: 10.1021/bi00186a019
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:66917
Collection:CaltechAUTHORS
Deposited By: Victoria Brennan
Deposited On:18 May 2016 23:30
Last Modified:09 Mar 2020 13:19

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