Post-translational modification of the deubiquitinating enzyme UCHL5 in the DNA damage response: mechanistic and preclinical approaches
The deubiquitinating enzyme (DUB) UCHL5 has been shown to have a role in the DNA damage response (DDR) and we have identified that UCHL5 is post-translationally acetylated in bladder cancer cells treated with histone deacetylase inhibitors (HDACi). HDAC inhibitors are known radiosensitisers with some systemic toxicities, but we have shown that they do not add to local normal tissue side effects of radiotherapy, thus increasing the radiotherapy therapeutic ratio. We wish to find similarly effective radiosensitisers which are less systemically toxic.
Our hypothesis is that UCHL5 acetylation is important for its function in the DDR, so that inhibition of UCHL5 or modulation of UCHL5 acetylation could be an effective and more targeted mechanism of radiosensitisation than HDACi.
In this project, the student shall characterise UCHL5 acetylation and its role in the DDR and establish whether chemical inhibitors of UCHL5 (DUBi) are effective alone or in combination with HDACi as radiosensitisers. Techniques will include standard molecular biology techniques, biochemical and biophysical assays, co-immunoprecipitation, clonogenic assays, live cell microscopy and in vitro and in vivo drug testing. We have also recently found that acetylation of multiple non-histone proteins is important in the DDR, so the student may study additional new targets. Secondary supervision will be provided by Kristijan Ramadan.