Brian Luke: Mechanistic understanding of Rtt101/CUL4 function at sites of single ribonucleotide insertions (Project 9, 2019-2022)
We aim at understanding the mechanisms that protect the genome from the adverse consequences of incorporating single ribonucleotides during DNA replication. To this end, we will investigate the activities of a conserved ubiquitin ligase, Cul4/Rtt101, at sites of ribonucleotide-induced replication stress, and determine its impact on replication dynamics by a combination of microscopy, chromatin immunoprecipitation and single DNA fibre analysis. We will characterise the DNA polymerase e subunit Dpb2 as a potential substrate of Cul4/Rtt101 in budding yeast and in the long term elucidate the role of human CUL4 in a genetic disease related to the phenomenon, Aicardi-Goutières syndrome.