FN Archimer Export Format
PT J
TI Calcium-driven DNA synthesis by a high-fidelity DNA polymerase
BT 
AF RALEC, Celine
   HENRY, Etienne
   LEMOR, Melanie
   KILLELEA, Tom
   HENNEKE, Ghislaine
AS 1:1,2,3;2:1,2,3;3:1,2,3;4:1,2,3;5:1,2,3;
FF 1:PDG-REM-EEP-LMEE;2:;3:;4:;5:PDG-REM-EEP-LMEE;
C1 IFREMER, Ctr Brest, LM2E, UMR 6197,Technopole Brest Iroise, F-29280 Plouzane, France.
   CNRS, LM2E, UMR 6197, Technopole Brest Iroise, F-29280 Plouzane, France.
   Univ Brest Occidentale, Technopole Brest Iroise, UMR 6197, LM2E, F-29280 Plouzane, France.
C2 IFREMER, FRANCE
   CNRS, FRANCE
   UBO, FRANCE
SI BREST
SE PDG-REM-EEP-LMEE
UM BEEP-LM2E
IN WOS Ifremer jusqu'en 2018
   DOAJ
   copubli-france
   copubli-univ-france
IF 11.561
TC 22
UR https://archimer.ifremer.fr/doc/00405/51663/52210.pdf
   https://archimer.ifremer.fr/doc/00405/51663/52211.pdf
LA English
DT Article
AB Divalent metal ions, usually Mg2+, are required for both DNA synthesis and proofreading functions by DNA polymerases (DNA Pol). Although used as a non-reactive cofactor substitute for binding and crystallographic studies, Ca2+ supports DNA polymerization by only one DNA Pol, Dpo4. Here, we explore whether Ca2+-driven catalysis might apply to high-fidelity (HiFi) family B DNA Pols. The consequences of replacing Mg2+ by Ca2+ on base pairing at the polymerase active site as well as the editing of terminal nucleotides at the exonuclease active site of the archaeal Pyrococcus abyssi DNA Pol (PabPolB) are characterized and compared to other (families B, A, Y, X, D) DNA Pols. Based on primer extension assays, steady-state kinetics and ion-chased experiments, we demonstrate that Ca2+ (and other metal ions) activates DNA synthesis by PabPolB. While showing a slower rate of phosphodiester bond formation, nucleotide selectivity is improved over that of Mg2+. Further mechanistic studies show that the affinities for primer/template are higher in the presence of Ca2+ and reinforced by a correct incoming nucleotide. Conversely, no exonuclease degradation of the terminal nucleotides occurs with Ca2+. Evolutionary and mechanistic insights among DNA Pols are thus discussed. 
PY 2017
PD DEC
SO Nucleic Acids Research
SN 0305-1048
PU Oxford Univ Press
VL 45
IS 21
UT 000417691300033
BP 12425
EP 12440
DI 10.1093/nar/gkx927
ID 51663
ER

EF