Translesion DNA Synthesis Across Lesions Induced by Oxidative Products of Pyrimidines: An Insight into the Mechanism by Microscale Thermophoresis

Autor: Viktor Brabec, Ondrej Hrabina, Olga Novakova
Rok vydání: 2019
Předmět:
0301 basic medicine
DNA Repair
DNA polymerase
2’-deoxyribo-5-hydroxyuridin
translesion dna synthesis
DNA-Directed DNA Polymerase
01 natural sciences
Pentoxyl
lcsh:Chemistry
chemistry.chemical_compound
lcsh:QH301-705.5
Spectroscopy
Klenow fragment
biology
food and beverages
General Medicine
Computer Science Applications
Biochemistry
Thermodynamics
Oxidation-Reduction
DNA Replication
Article
Catalysis
Inorganic Chemistry
03 medical and health sciences
Humans
Physical and Theoretical Chemistry
Uracil
2’-deoxyribo-5-hydroxymethyl- uridin
oxidized nucleotides
Molecular Biology
DNA synthesis
010405 organic chemistry
Microscale thermophoresis
fungi
Organic Chemistry
dna polymerases
DNA
microscale thermophoresis
Reverse transcriptase
0104 chemical sciences
Oxidative Stress
Pyrimidines
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
chemistry
HIV-1
biology.protein
DNA polymerase I
Thymidine
DNA Damage
Mutagens
Zdroj: International Journal of Molecular Sciences, Vol 20, Iss 20, p 5012 (2019)
International Journal of Molecular Sciences
Volume 20
Issue 20
ISSN: 1422-0067
DOI: 10.3390/ijms20205012
Popis: Oxidative stress in cells can lead to the accumulation of reactive oxygen species and oxidation of DNA precursors. Oxidized nucleotides such as 2&rsquo
deoxyribo-5-hydroxyuridin (HdU) and 2&rsquo
deoxyribo-5-hydroxymethyluridin (HMdU) can be inserted into DNA during replication and repair. HdU and HMdU have attracted particular interest because they have different effects on damaged-DNA processing enzymes that control the downstream effects of the lesions. Herein, we studied the chemically simulated translesion DNA synthesis (TLS) across the lesions formed by HdU or HMdU using microscale thermophoresis (MST). The thermodynamic changes associated with replication across HdU or HMdU show that the HdU paired with the mismatched deoxyribonucleoside triphosphates disturbs DNA duplexes considerably less than thymidine (dT) or HMdU. Moreover, we also demonstrate that TLS by DNA polymerases across the lesion derived from HdU was markedly less extensive and potentially more mutagenic than that across the lesion formed by HMdU. Thus, DNA polymerization by DNA polymerase &eta
(pol&eta
), the exonuclease-deficient Klenow fragment of DNA polymerase I (KF&ndash
), and reverse transcriptase from human immunodeficiency virus type 1 (HIV-1 RT) across these pyrimidine lesions correlated with the different stabilization effects of the HdU and HMdU in DNA duplexes revealed by MST. The equilibrium thermodynamic data obtained by MST can explain the influence of the thermodynamic alterations on the ability of DNA polymerases to bypass lesions induced by oxidative products of pyrimidines. The results also highlighted the usefulness of MST in evaluating the impact of oxidative products of pyrimidines on the processing of these lesions by damaged DNA processing enzymes.
Databáze: OpenAIRE
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