MS Albelazi
The biochemical role of the human NEIL1 and NEIL3 DNA glycosylases on model DNA replication forks
Albelazi, MS; Martin, PR; Mohammed, S; Mutti, L; Parsons, JL; Elder, RH
Authors
PR Martin
S Mohammed
L Mutti
JL Parsons
RH Elder
Abstract
Endonuclease VIII-like (NEIL) 1 and 3 proteins eliminate oxidative DNA base damage and psoralen DNA interstrand crosslinks through initiation of base excision repair. Current evidence points to a DNA replication associated repair function of NEIL1 and NEIL3, correlating with induced expression of the proteins in S/G2 phases of the cell cycle. However previous attempts to express and purify recombinant human NEIL3 in an active form have been challenging. In this study, both human NEIL1 and NEIL3 have been expressed and purified from E. coli, and the DNA glycosylase activity of these two proteins confirmed using single- and double-stranded DNA oligonucleotide substrates containing the oxidative bases, 5-hydroxyuracil, 8-oxoguanine and thymine glycol. To determine the biochemical role that NEIL1 and NEIL3 play during DNA replication, model replication fork substrates were designed containing the oxidized bases at one of three specific sites relative to the fork. Results indicate that whilst specificity for 5- hydroxyuracil and thymine glycol was observed, NEIL1 acts preferentially on double-stranded DNA, including the damage upstream to the replication fork, whereas NEIL3 preferentially excises oxidized bases from single stranded DNA and within open fork structures. Thus, NEIL1 and NEIL3 act in concert to remove oxidized bases from the replication fork.
Keywords: base excision repair; DNA damage; DNA repair; DNA replication; NEI-like DNA glycosylases
Citation
Albelazi, M., Martin, P., Mohammed, S., Mutti, L., Parsons, J., & Elder, R. (2019). The biochemical role of the human NEIL1 and NEIL3 DNA glycosylases on model DNA replication forks. Genes, 10(4), 315. https://doi.org/10.3390/genes10040315
Journal Article Type | Article |
---|---|
Acceptance Date | Apr 17, 2019 |
Online Publication Date | Apr 23, 2019 |
Publication Date | Apr 23, 2019 |
Deposit Date | Apr 23, 2019 |
Publicly Available Date | Apr 23, 2019 |
Journal | Genes |
Publisher | MDPI |
Volume | 10 |
Issue | 4 |
Pages | 315 |
DOI | https://doi.org/10.3390/genes10040315 |
Publisher URL | https://doi.org/10.3390/genes10040315 |
Related Public URLs | https://www.mdpi.com/journal/genes |
Files
genes-10-00315.pdf
(1.3 Mb)
PDF
Licence
http://creativecommons.org/licenses/by/4.0/
Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/
Downloadable Citations
About USIR
Administrator e-mail: library-research@salford.ac.uk
This application uses the following open-source libraries:
SheetJS Community Edition
Apache License Version 2.0 (http://www.apache.org/licenses/)
PDF.js
Apache License Version 2.0 (http://www.apache.org/licenses/)
Font Awesome
SIL OFL 1.1 (http://scripts.sil.org/OFL)
MIT License (http://opensource.org/licenses/mit-license.html)
CC BY 3.0 ( http://creativecommons.org/licenses/by/3.0/)
Powered by Worktribe © 2024
Advanced Search