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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

The biochemical role of the human NEIL1 and NEIL3 DNA glycosylases on model DNA replication forks Thumbnail


Authors

MS Albelazi

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

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