Reading ADP-ribosylation signalling using chemical biology and interaction proteomics
Description
ADP-ribose (ADPr) readers are essential components of ADP-ribosylation signalling, which regulates genome maintenance and immunity. The identification and discrimination between monoADPr (MAR) and polyADPr (PAR) readers is difficult due to a lack of suitable affinity enrichment reagents. We synthesized well-defined ADPr probes and used these for affinity purifications combined with relative and absolute quantitative mass spectrometry to generate proteome-wide MAR and PAR interactomes, including determination of apparent binding affinities. Amongst the main findings, MAR and PAR readers regulate various common and distinct processes, such as the DNA damage response, cellular metabolism, RNA trafficking and transcription. We monitored the dynamics of PAR interactions upon induction of oxidative DNA damage and uncovered new mechanistic connections between ubiquitin signalling and ADP-ribosylation. Taken together, chemical biology enables exploration of MAR and PAR readers using interaction proteomics. Furthermore, the generated MAR and PAR interaction maps significantly expand our current understanding of ADPr signalling. The files contain source data for WB images.
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Steps to reproduce
Detailed information about WB can be found in the publication.