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A single N -2-acetylaminofluorene adduct alters the footprint of T7 (exo−) DNA polymerase bound to a model primer–template junction

Identifieur interne : 003B00 ( Main/Exploration ); précédent : 003A99; suivant : 003B01

A single N -2-acetylaminofluorene adduct alters the footprint of T7 (exo−) DNA polymerase bound to a model primer–template junction

Auteurs : Dominique Y. Burnouf [France] ; Robert P. P Fuchs [France]

Source :

Mutation Research-DNA Repair [ 0921-8777 ] ; 1998.

RBID : ISTEX:41F5B8C605DEE736A1A79206463C18CF321E641F

English descriptors

Teeft :
- Acad, Adduct, Base pairs, Biochemistry, Biol, Burnouf, Chem, Coli, Conformation, Covalent binding, Digestion, Dnasei, Dnasei cleavage, Dnasei digestion, Duplex, Escherichia, Escherichia coli, Exonuclease activity, Final concentration, Footprint, Footprint analysis, Footprinting, Fuchs, Fuchsr, Fuchsr mutation research, Guanine, Helix, Holoenzyme, Hypersensitive, Hypersensitive sites, Kinetic experiments, Klenow fragment, Lesion, Model junction, Molecular dynamics, Monomodified, Monomodified duplex, Mutation, Natl, Nucleotide, Oligonucleotide, Phosphodiester, Phosphodiester bonds, Phosphodiester bounds, Polymerase, Polymerase binding, Polymerase cleft, Polymerase holoenzyme, Primer, Proc, Replication, Same position, Template strand, Unmodified, Unmodified duplex, Unmodified duplexes.

Abstract

Abstract: Bovine pancreatic deoxyribonuclease I (DNaseI) has been used to footprint T7 (exo−) DNA polymerase bound to a model primer–template junction. The polymerase was blocked at a specific position either by the omission of dCTP from the reaction mix or by the presence of a N-(deoxyguanosin-8-yl)-2-acetylaminofluorene (dGuo-AAF) adduct. This lesion has been shown to be a severe block for several DNA polymerases, both in in vitro primer elongation experiments, and during the in vivo replication of AAF-monomodified single-stranded vectors. The footprints obtained with unmodified primer–template DNA define two protected domains separated by an inter-region that remains sensitive to DNaseI, and several hypersensitive sites located on both strands. Binding of the polymerase to AAF monomodified duplexes results in the same protection pattern as that obtained with the unmodified duplexes. However, the hypersensitive sites either disappear or are dramatically reduced. The results suggest that the AAF lesion alters the correct positioning of the duplex DNA within the polymerase cleft.

Url:

https://api.istex.fr/ark:/67375/6H6-FMNSS9ST-4/fulltext.pdf

DOI: 10.1016/S0921-8777(97)00058-X

Affiliations:

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Le document en format XML

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<term>Digestion</term>
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<term>Footprinting</term>
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<front><div type="abstract" xml:lang="en">Abstract: Bovine pancreatic deoxyribonuclease I (DNaseI) has been used to footprint T7 (exo−) DNA polymerase bound to a model primer–template junction. The polymerase was blocked at a specific position either by the omission of dCTP from the reaction mix or by the presence of a N-(deoxyguanosin-8-yl)-2-acetylaminofluorene (dGuo-AAF) adduct. This lesion has been shown to be a severe block for several DNA polymerases, both in in vitro primer elongation experiments, and during the in vivo replication of AAF-monomodified single-stranded vectors. The footprints obtained with unmodified primer–template DNA define two protected domains separated by an inter-region that remains sensitive to DNaseI, and several hypersensitive sites located on both strands. Binding of the polymerase to AAF monomodified duplexes results in the same protection pattern as that obtained with the unmodified duplexes. However, the hypersensitive sites either disappear or are dramatically reduced. The results suggest that the AAF lesion alters the correct positioning of the duplex DNA within the polymerase cleft.</div>
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A single N -2-acetylaminofluorene adduct alters the footprint of T7 (exo−) DNA polymerase bound to a model primer–template junction

A single N -2-acetylaminofluorene adduct alters the footprint of T7 (exo−) DNA polymerase bound to a model primer–template junction

Source :

English descriptors

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri