UMMS Affiliation

Department of Biochemistry and Molecular Pharmacology

Publication Date

1-25-1994

Document Type

Article

Subjects

Adenine Nucleotides; Aphidicolin; Bacteriophage T4; Base Sequence; DNA-Directed DNA Polymerase; Deoxyguanine Nucleotides; Guanine Nucleotides; Kinetics; Molecular Sequence Data

Disciplines

Biochemistry

Abstract

Bacteriophage T4 DNA polymerase was inhibited by butylphenyl nucleotides, aphidicolin and pyrophosphate analogs, but with lower sensitivities than other members of the B family DNA polymerases. The nucleotides N2-(p-n-butylphenyl)dGTP (BuPdGTP) and 2-(p-n-butylanilino)dATP (BuAdATP) inhibited T4 DNA polymerase with competitive Ki values of 0.82 and 0.54 microM with respect to dGTP and dATP, respectively. The same compounds were more potent inhibitors in truncated assays lacking the competitor dNTP, displaying apparent Ki values of 0.001 and 0.0016 microM, respectively. BuPdGTP was a substrate for T4 DNA polymerase, and the resulting 3'-BuPdG-primer:template was bound strongly by the enzyme. Each of the non-substrate derivatives, BuPdGDP and BuPdGMPCH2PP, inhibited T4 DNA polymerase with similar potencies in both the truncated and variable competitor assays. These results indicate that BuPdGTP inhibits T4 DNA polymerase by distinct mechanisms depending upon the assay conditions. Reversible competitive inhibition predominates in the presence of dGTP, and incorporation in the absence of dGTP leads to potent inhibition by the modified primer:template. The implications of these findings for the use of these inhibitors in the study of B family DNA polymerases is discussed.

Rights and Permissions

Citation: Nucleic Acids Res. 1994 Jan 25;22(2):232-7. doi: 10.1093/nar/22.2.232. Link to article on publisher's website

DOI of Published Version

10.1093/nar/22.2.232

Comments

Publisher PDF posted as allowed by the publisher's author rights policy at http://www.jneurosci.org/site/misc/ifa_policies.xhtml#copyright.

Related Resources

Link to Article in PubMed

Journal/Book/Conference Title

Nucleic acids research

PubMed ID

8121808

Included in

Biochemistry Commons

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.