Biology subjects

Drake, M.

Publications and source records attributed to Drake, M..

2 recordsLinked to original sources

bioRxiv2023

The RPA complex orchestrates K63-linked deubiquitination via ZUP1

Ubiquitin signaling is regulated by deubiquitinating enzymes (DUBs), which can edit or remove ubiquitin from substrates. Recently, ZUP1 was discovered as the founding and only member of a novel DUB class that cleaves long K63-linked ubiquitin chains, with a putative role in DNA repair. However, ZUP1 has poor activity on its own, suggesting additional mechanisms exist to promote its activity in cells. Here, using a range of cellular, biochemical, and structural proteomics approaches, we show that ZUP1 directly interacts with the RPA complex, a single-stranded DNA binding protein complex involved in DNA replication and multiple DNA repair processes. Functionally, the ZUP1-RPA complex interaction dramatically stimulates ZUP1 K63-linked DUB activity, which occurs through S1 and S1 site communication. Collectively, our results suggest a mechanism that couples sensing of ssDNA to the activation of K63-linked deubiquitination via the ZUP1-RPA complex.

biochemistry↗

bioRxiv2021

Collateral damage: Identification and characterisation of spontaneous mutations causing deafness from a targeted knockout programme

Mice carrying targeted mutations are important for investigating gene function and the role of genes in disease, but the process of culturing embryonic stem cells during the making of a targeted allele offers opportunities for spontaneous mutations to arise. Identifying spontaneous mutations relies on the detection of phenotypes segregating independently of targeted alleles, and many phenotypes are easy to miss if not specifically looked for. Here we present data from a large, targeted knockout programme in which mice were analysed through a phenotyping pipeline. Twenty-five lines out of 1311 displayed different deafness phenotypes that did not segregate with the targeted allele. We have identified 8 different mutations causing deafness in 16 of these 25 lines and characterised the resulting phenotypes. Our data show that spontaneous mutations with observable effects on phenotype are a common side effect of intensive breeding programmes, including those underlying targeted mutation programmes.

genetics↗