Ubiquitylation-dependent regulation of NEIL1 by Mule and TRIM26 is required for the cellular DNA damage response

Edmonds, MJ, Carter, RJ, Nickson, CM ORCID: 0000-0003-4611-2852, Williams, SC and Parsons, JL ORCID: 0000-0002-5052-1125 (2017) Ubiquitylation-dependent regulation of NEIL1 by Mule and TRIM26 is required for the cellular DNA damage response. Nucleic Acids Research, 45 (2). pp. 726-738.

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Abstract

Endonuclease VIII-like protein 1 (NEIL1) is a DNA glycosylase involved in initiating the base excision repair pathway, the major cellular mechanism for repairing DNA base damage. Here, we have purified the major E3 ubiquitin ligases from human cells responsible for regulation of NEIL1 by ubiquitylation. Interestingly, we have identified two enzymes that catalyse NEIL1 polyubiquitylation, Mcl-1 ubiquitin ligase E3 (Mule) and tripartite motif 26 (TRIM26). We demonstrate that these enzymes are capable of polyubiquitylating NEIL1 in vitro, and that both catalyse ubiquitylation of NEIL1 within the same C-terminal lysine residues. An siRNA-mediated knockdown of Mule or TRIM26 leads to stabilisation of NEIL1, demonstrating that these enzymes are important in regulating cellular NEIL1 steady state protein levels. Similarly, a mutant NEIL1 protein lacking residues for ubiquitylation is more stable than the wild type protein in vivo. We also demonstrate that cellular NEIL1 protein is induced in response to ionising radiation (IR), although this occurs specifically in a Mule-dependent manner. Finally we show that stabilisation of NEIL1, particularly following TRIM26 siRNA, contributes to cellular resistance to IR. This highlights the importance of Mule and TRIM26 in maintaining steady state levels of NEIL1, but also those required for the cellular DNA damage response.

Item Type:	Article
Uncontrolled Keywords:	Hela Cells, Humans, DNA Damage, DNA Glycosylases, Ubiquitin-Protein Ligases, DNA-Binding Proteins, Tumor Suppressor Proteins, Recombinant Fusion Proteins, DNA Repair, Gene Expression Regulation, Protein Binding, Radiation, Ionizing, Radiation Tolerance, Ubiquitination, Tripartite Motif Proteins
Depositing User:	Symplectic Admin
Date Deposited:	03 Nov 2016 14:23
Last Modified:	19 Jan 2023 07:26
DOI:	10.1093/nar/gkw959
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3004326