TY - JOUR
T1 - Novel inhibition of archaeal family-D DNA polymerase by uracil
AU - Richardson, Tomas T.
AU - Gilroy, Louise
AU - Ishino, Yoshizumi
AU - Connolly, Bernard A.
AU - Henneke, Ghislaine
N1 - Funding Information:
French National Research Agency [ANR-10-JCJC-1501-01 to G.H.]; T.T.R. and L.G. are UK BBSRC supported PhD students. Funding for open access charge: French National Research Agency (ANR).
PY - 2013/4
Y1 - 2013/4
N2 - Archaeal family-D DNA polymerase is inhibited by the presence of uracil in DNA template strands. When the enzyme encounters uracil, following three parameters change: DNA binding increases roughly 2-fold, the rate of polymerization slows by a factor of ∼5 and 3′-5′ proof-reading exonuclease activity is stimulated by a factor of ∼2. Together these changes result in a significant decrease in polymerization activity and a reduction in net DNA synthesis. Pol D appears to interact with template strand uracil irrespective of its distance ahead of the replication fork. Polymerization does not stop at a defined location relative to uracil, rather a general decrease in DNA synthesis is observed. 'Trans' inhibition, the slowing of Pol D by uracil on a DNA strand not being replicated is also observed. It is proposed that Pol D is able to interact with uracil by looping out the singlestranded template, allowing simultaneous contact of both the base and the primer-template junction to give a polymerase-DNA complex with diminished extension ability.
AB - Archaeal family-D DNA polymerase is inhibited by the presence of uracil in DNA template strands. When the enzyme encounters uracil, following three parameters change: DNA binding increases roughly 2-fold, the rate of polymerization slows by a factor of ∼5 and 3′-5′ proof-reading exonuclease activity is stimulated by a factor of ∼2. Together these changes result in a significant decrease in polymerization activity and a reduction in net DNA synthesis. Pol D appears to interact with template strand uracil irrespective of its distance ahead of the replication fork. Polymerization does not stop at a defined location relative to uracil, rather a general decrease in DNA synthesis is observed. 'Trans' inhibition, the slowing of Pol D by uracil on a DNA strand not being replicated is also observed. It is proposed that Pol D is able to interact with uracil by looping out the singlestranded template, allowing simultaneous contact of both the base and the primer-template junction to give a polymerase-DNA complex with diminished extension ability.
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U2 - 10.1093/nar/gkt083
DO - 10.1093/nar/gkt083
M3 - Article
C2 - 23408858
AN - SCOPUS:84876573014
SN - 0305-1048
VL - 41
SP - 4207
EP - 4218
JO - Nucleic Acids Research
JF - Nucleic Acids Research
IS - 7
ER -