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Modulation of the Apurinic/Apyrimidinic Endonuclease Activity of Human APE1 and of Its Natural Polymorphic Variants by Base Excision Repair Proteins.
- Source :
-
International journal of molecular sciences [Int J Mol Sci] 2020 Sep 28; Vol. 21 (19). Date of Electronic Publication: 2020 Sep 28. - Publication Year :
- 2020
-
Abstract
- Human apurinic/apyrimidinic endonuclease 1 (APE1) is known to be a critical player of the base excision repair (BER) pathway. In general, BER involves consecutive actions of DNA glycosylases, AP endonucleases, DNA polymerases, and DNA ligases. It is known that these proteins interact with APE1 either at upstream or downstream steps of BER. Therefore, we may propose that even a minor disturbance of protein-protein interactions on the DNA template reduces coordination and repair efficiency. Here, the ability of various human DNA repair enzymes (such as DNA glycosylases OGG1, UNG2, and AAG; DNA polymerase PolĪ²; or accessory proteins XRCC1 and PCNA) to influence the activity of wild-type (WT) APE1 and its seven natural polymorphic variants (R221C, N222H, R237A, G241R, M270T, R274Q, and P311S) was tested. Förster resonance energy transfer-based kinetic analysis of abasic site cleavage in a model DNA substrate was conducted to detect the effects of interacting proteins on the activity of WT APE1 and its single-nucleotide polymorphism (SNP) variants. The results revealed that WT APE1 activity was stimulated by almost all tested DNA repair proteins. For the SNP variants, the matters were more complicated. Analysis of two SNP variants, R237A and G241R, suggested that a positive charge in this area of the APE1 surface impairs the protein-protein interactions. In contrast, variant R221C (where the affected residue is located near the DNA-binding site) showed permanently lower activation relative to WT APE1, whereas neighboring SNP N222H did not cause a noticeable difference as compared to WT APE1. Buried substitution P311S had an inconsistent effect, whereas each substitution at the DNA-binding site, M270T and R274Q, resulted in the lowest stimulation by BER proteins. Protein-protein molecular docking was performed between repair proteins to identify amino acid residues involved in their interactions. The data uncovered differences in the effects of BER proteins on APE1, indicating an important role of protein-protein interactions in the coordination of the repair pathway.
- Subjects :
- Amino Acid Substitution
Binding Sites
DNA genetics
DNA metabolism
DNA Damage
DNA Glycosylases genetics
DNA Glycosylases metabolism
DNA Polymerase beta chemistry
DNA Polymerase beta genetics
DNA Polymerase beta metabolism
DNA-(Apurinic or Apyrimidinic Site) Lyase genetics
DNA-(Apurinic or Apyrimidinic Site) Lyase metabolism
Gene Expression
Humans
Kinetics
Molecular Docking Simulation
Nucleic Acid Conformation
Oligodeoxyribonucleotides chemistry
Oligodeoxyribonucleotides genetics
Oligodeoxyribonucleotides metabolism
Polymorphism, Single Nucleotide
Proliferating Cell Nuclear Antigen chemistry
Proliferating Cell Nuclear Antigen genetics
Proliferating Cell Nuclear Antigen metabolism
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Recombinant Proteins chemistry
Recombinant Proteins genetics
Recombinant Proteins metabolism
Substrate Specificity
X-ray Repair Cross Complementing Protein 1 chemistry
X-ray Repair Cross Complementing Protein 1 genetics
X-ray Repair Cross Complementing Protein 1 metabolism
DNA chemistry
DNA Glycosylases chemistry
DNA Repair
DNA-(Apurinic or Apyrimidinic Site) Lyase chemistry
Subjects
Details
- Language :
- English
- ISSN :
- 1422-0067
- Volume :
- 21
- Issue :
- 19
- Database :
- MEDLINE
- Journal :
- International journal of molecular sciences
- Publication Type :
- Academic Journal
- Accession number :
- 32998246
- Full Text :
- https://doi.org/10.3390/ijms21197147