Efficient DNA damage response (DDR) is a fundamental process for every living organism. The accumulation of DNA damage affects cell viability and leads to a variety of diseases, particularly cancer. Therefore, understanding of the molecular mechanisms necessary for DNA damage repair is of great importance. A myriad of repair factors targets lethal double-strand breaks (DSBs) by non-homologous end joining (NHEJ) and homologous recombination (HR) pathways. 53BP1 is a key regulator of DSB repair pathway choice. It promotes (NHEJ)-mediated DSB repair by antagonizing long-range DNA end-resection, which is essential for DSB repair via homologous recombination.
TIRR/NUDT16L1 is a recently identified binding partner and negative regulator of 53BP1 (Drané et al., 2017; Dai et al., 2018; Wang et al., 2018). In non-damage conditions, TIRR and 53BP1 are bound. However, upon the occurrence of DNA double strand breaks (DSBs), TIRR and 53BP1 separate and 53BP1 binds the chromatin at the break site (Drané et al., 2017).
Currently, very little is known about TIRR and its roles within cells both in damage and non-damage contexts. Mass spectrometry to identify TIRR interacting partners has been carried out only in non-damage conditions (Drané et al., 2017; Avolio et al., 2018). We are therefore in the process of establishing a proximity labelling BioID technique on TIRR in both non-damage and damage conditions to characterise the interactome of TIRR and to identify novel roles of TIRR. Additionally, we hope to integrate this BioID data with existing data generated in TIRR knockdown conditions. More than 10% of cancer cell lines are highly sensitive to TIRR knockdown. We hope to identify what underlines this phenotype and how this could be used for development of novel cancer therapy.
Therefore, the main aims of this project would be:
- To use data generated from this BioID approach to identify and validate novel TIRR interactors in damage and non-damage conditions.
- Use Go analysis of BioID data and integrate this with other existing data to identify novel roles of TIRR in cells.
- Use functional assays including survival assays to investigate cellular phenotypes in TIRR knockdown conditions.
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