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UMass Chan Affiliations
Department of Biochemistry and Molecular PharmacologyDocument Type
Journal ArticlePublication Date
1994-01-25Keywords
Adenine NucleotidesAphidicolin
Bacteriophage T4
Base Sequence
DNA-Directed DNA Polymerase
Deoxyguanine Nucleotides
Guanine Nucleotides
Kinetics
Molecular Sequence Data
Biochemistry
Metadata
Show full item recordAbstract
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.Source
Nucleic Acids Res. 1994 Jan 25;22(2):232-7. doi: 10.1093/nar/22.2.232. Link to article on publisher's websiteDOI
10.1093/nar/22.2.232Permanent Link to this Item
http://hdl.handle.net/20.500.14038/38870PubMed ID
8121808Related Resources
Link to Article in PubMedRights
Publisher PDF posted as allowed by the publisher's author rights policy at http://www.jneurosci.org/site/misc/ifa_policies.xhtml#copyright.
ae974a485f413a2113503eed53cd6c53
10.1093/nar/22.2.232