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Repair activity of the polymorphic Cys 326 variant of OGG1 under conditions of oxidative stress

Kaur, Mehak Preet (2016)
Other thesis, University of Birmingham.

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Reactive oxygen species (ROS) have damaging effect on cell metabolism and function via various DNA and protein modifications. Cells have evolved many mechanisms to reduce such ROS-induced damage, for instance the base excision repair (BER) pathway. This research emphases the effect of the common Ser326Cys polymorphism in the OGG 1 gene, which is an important part of BER to remove mutagenic 8-oxo deoxyguanosine from genomic DNA. Previous studies have highlighted the importance of Ser326Cys polymorphism of OGG 1 relating it to many forms of cancer and various degenerative diseases due to variant OGG 1 been reported being repair deficient. In the current study, we report the novel application of bimolecular
fluorescence complementation (BiFC) to study OGG 1 protein complex formation in the native cellular environment and report accumulation of Cys326-0GG 1 protein complexes as assessed by confocal microscopy. We observed fluorescence both in and around the nucleus of cells expressing Cys326-0GG 1 specifically under conditions of cellular oxidative stress, with increased ROS levels and depletion of GSH following treatment with both buthionine sulfoximine (BSO) and hydrogen peroxide. Furthermore, loss of BiFC fluorescence was seen when cells were treated with thiol reducing agents providing evidence of the role a disulfide bond. In summary, we report for the first time formation of Cys326 OGG 1 complexes in the native cellular environment particularly under conditions of oxidative stress. This finding could
explain the reduced repair capability of Cys3260GG 1 under oxidative stress environment.

Type of Work:MSc by Res thesis.
Supervisor(s):Hodges, Nik
School/Faculty:Colleges (2008 onwards) > College of Life & Environmental Sciences
Department:School of Biosciences
Subjects:QR Microbiology
Institution:University of Birmingham
ID Code:6853
This unpublished thesis/dissertation is copyright of the author and/or third parties. The intellectual property rights of the author or third parties in respect of this work are as defined by The Copyright Designs and Patents Act 1988 or as modified by any successor legislation. Any use made of information contained in this thesis/dissertation must be in accordance with that legislation and must be properly acknowledged. Further distribution or reproduction in any format is prohibited without the permission of the copyright holder.
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