Translation-coupled sensing and degradation of RNA-protein crosslinks

Reactive aldehydes are abundant cytotoxic metabolites, which challenge homoeostasis by crosslinking cellular macromolecules. Whether RNA damage contributes to the toxicity of aldehydes and whether cells possess mechanisms to resolve RNA-protein crosslinks (RPCs) in particular is unknown. Studying the specific consequences of aldehyde-induced RNA damage is challenging due to confounding induction of DNA damage. Here, we establish photoactivatable ribonucleosides as a tractable model system to study aldehyde-mimicking RNA damage in the absence of DNA damage. The aim of this phosphoproteome measurement is to elucidate the changes in phosphorylation sites upon RPC induction using the treatment with a photoactivatable-ribonucleoside-enhanced crosslinking (PAR-CL) by incubation with 4-thiouridine (4-SU) and subsequent crosslinking by UVA irradiation.