A sensor complements the steric gate when DNA polymerase ϵ discriminates ribonucleotides
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A sensor complements the steric gate when DNA polymerase ϵ discriminates ribonucleotides. / Parkash, Vimal; Kulkarni, Yashraj; Bylund, Göran O.; Osterman, Pia; Kamerlin, Shina Caroline Lynn; Johansson, Erik.
I: Nucleic Acids Research, Bind 51, Nr. 20, 2023, s. 11225-11238.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - A sensor complements the steric gate when DNA polymerase ϵ discriminates ribonucleotides
AU - Parkash, Vimal
AU - Kulkarni, Yashraj
AU - Bylund, Göran O.
AU - Osterman, Pia
AU - Kamerlin, Shina Caroline Lynn
AU - Johansson, Erik
N1 - Publisher Copyright: © 2023 The Author(s). Published by Oxford University Press on behalf of Nucleic Acids Research.
PY - 2023
Y1 - 2023
N2 - The cellular imbalance between high concentrations of ribonucleotides (NTPs) and low concentrations of deoxyribonucleotides (dNTPs), is challenging for DNA polymerases when building DNA from dNTPs. It is currently believed that DNA polymerases discriminate against NTPs through a steric gate model involving a clash between a tyrosine and the 2′-hydroxyl of the ribonucleotide in the polymerase active site in B-family DNA polymerases. With the help of crystal structures of a B-family polymerase with a UTP or CTP in the active site, molecular dynamics simulations, biochemical assays and yeast genetics, we have identified a mechanism by which the finger domain of the polymerase sense NTPs in the polymerase active site. In contrast to the previously proposed polar filter, our experiments suggest that the amino acid residue in the finger domain senses ribonucleotides by steric hindrance. Furthermore, our results demonstrate that the steric gate in the palm domain and the sensor in the finger domain are both important when discriminating NTPs. Structural comparisons reveal that the sensor residue is conserved among B-family polymerases and we hypothesize that a sensor in the finger domain should be considered in all types of DNA polymerases.
AB - The cellular imbalance between high concentrations of ribonucleotides (NTPs) and low concentrations of deoxyribonucleotides (dNTPs), is challenging for DNA polymerases when building DNA from dNTPs. It is currently believed that DNA polymerases discriminate against NTPs through a steric gate model involving a clash between a tyrosine and the 2′-hydroxyl of the ribonucleotide in the polymerase active site in B-family DNA polymerases. With the help of crystal structures of a B-family polymerase with a UTP or CTP in the active site, molecular dynamics simulations, biochemical assays and yeast genetics, we have identified a mechanism by which the finger domain of the polymerase sense NTPs in the polymerase active site. In contrast to the previously proposed polar filter, our experiments suggest that the amino acid residue in the finger domain senses ribonucleotides by steric hindrance. Furthermore, our results demonstrate that the steric gate in the palm domain and the sensor in the finger domain are both important when discriminating NTPs. Structural comparisons reveal that the sensor residue is conserved among B-family polymerases and we hypothesize that a sensor in the finger domain should be considered in all types of DNA polymerases.
U2 - 10.1093/nar/gkad817
DO - 10.1093/nar/gkad817
M3 - Journal article
C2 - 37819038
AN - SCOPUS:85178042069
VL - 51
SP - 11225
EP - 11238
JO - Nucleic Acids Research
JF - Nucleic Acids Research
SN - 0305-1048
IS - 20
ER -
ID: 375589495