Identifying the protein interactions of Mre11-Rad50 in Haloferax volcanii during double-strand break repair

Wickham-Smith, Charlie (2015) Identifying the protein interactions of Mre11-Rad50 in Haloferax volcanii during double-strand break repair. MRes thesis, University of Nottingham.

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The Mre11 and Rad50 proteins are found in all domains of life and form a complex that locates and binds to DNA double-strand breaks (DSBs). The complex tethers DNA ends and coordinates the repair of DSBs. In Haloferax volcanii, mre11rad50 mutants are more resistant to DNA damage than the wild-type. Mre11-Rad50 is believed to restrain repair of DSBs by homologous recombination (HR) while other repair pathways can operate. Mutants of mre11rad50 cannot utilise this restraining mechanism so HR is believed to solely repair DSBs, leading to an increase in DNA damage resistance. H. volcanii is highly polyploid, possessing up to 20 copies of its genome. This restraining method of repair will prevent DNA ends engaging with multiple partners, which could be highly toxic. The specific details of how H. volcanii repair DSBs are unknown, including the protein interaction of Mre11-Rad50.

Mre11-Rad50 has been labelled with His6 and StrepII tags then purified on gravity columns. Several proteins have been shown to co-purify with His6/StrepII-tagged Mre11-Rad50 under normal growth conditions. To discover whether these proteins are present during DSB repair, protein purification following incubation with DNA damaging agents is required. Further analysis of these proteins will reveal their identity and whether they are playing a role in DSB repair in H. volcanii

Item Type: Thesis (University of Nottingham only) (MRes)
Supervisors: Allers, Thorsten
Subjects: Q Science > QH Natural history. Biology > QH426 Genetics
Faculties/Schools: UK Campuses > Faculty of Medicine and Health Sciences > School of Life Sciences
Item ID: 30816
Depositing User: Wickham-Smith, Charles
Date Deposited: 10 Aug 2016 08:20
Last Modified: 14 Sep 2016 05:43

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