NTH1 is a new target for ubiquitylation-dependent regulation by TRIM26 required for the cellular response to oxidative stress



Williams, Sarah C and Parsons, Jason L ORCID: 0000-0002-5052-1125
(2018) NTH1 is a new target for ubiquitylation-dependent regulation by TRIM26 required for the cellular response to oxidative stress. Molecular and cellular biology, 38 (12).

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Abstract

Endonuclease III-like protein 1 (NTH1) is a DNA glycosylase required for the repair of oxidised bases, such as thymine glycol, during the base excision repair pathway. We examined regulation of NTH1 protein by the ubiquitin proteasome pathway and have identified the E3 ubiquitin ligase tripartite motif 26 (TRIM26) as the major enzyme targeting NTH1 for polyubiquitylation. We demonstrate that TRIM26 catalyses ubiquitylation of NTH1 predominantly on lysine 67 present within the N-terminus of the protein in vitro In addition, the stability of a ubiquitylation-deficient protein mutant of NTH1 (lysine to arginine) at this specific residue is significantly increased in comparison to the wild type protein when transiently expressed in cultured cells. We also demonstrate that cellular NTH1 protein is induced in response to oxidative stress following hydrogen peroxide treatment of cells, and that accumulation of NTH1 on chromatin is exacerbated in the absence of TRIM26 through siRNA depletion. Stabilisation of NTH1 following TRIM26 siRNA also causes a significant acceleration in the kinetics of DNA damage repair and cellular resistance to oxidative stress, which can be recapitulated by overexpression of NTH1. This demonstrates the importance of TRIM26 in regulating the cellular levels of NTH1, particularly under conditions of oxidative stress.

Item Type: Article
Uncontrolled Keywords: DNA damage, DNA glycosylase, DNA repair, base excision repair, oxidative stress, ubiquitination
Depositing User: Symplectic Admin
Date Deposited: 04 May 2018 08:32
Last Modified: 19 Jan 2023 06:34
DOI: 10.1128/mcb.00616-17
Related URLs:
URI: https://livrepository.liverpool.ac.uk/id/eprint/3020896