Comprehensive ADP‐ribosylome analysis identifies tyrosine as an ADP‐ribose acceptor site |
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| Authors: | Vera Bilan Kathrin Nowak Kapila Gunasekera Elena Ferrari Ralph Imhof Lars Malmström Michael O Hottiger |
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| Affiliation: | 1. Department of Molecular Mechanisms of Disease, University of Zurich, Zurich, Switzerland;2. Molecular Life Science PhD Program of the Life Science Zurich Graduate School, Zurich, Switzerland;3. S3IT and Institute for Computational Science, University of Zurich, Zurich, Switzerland |
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| Abstract: | Despite recent mass spectrometry (MS)‐based breakthroughs, comprehensive ADP‐ribose (ADPr)‐acceptor amino acid identification and ADPr‐site localization remain challenging. Here, we report the establishment of an unbiased, multistep ADP‐ribosylome data analysis workflow that led to the identification of tyrosine as a novel ARTD1/PARP1‐dependent in vivo ADPr‐acceptor amino acid. MS analyses of in vitro ADP‐ribosylated proteins confirmed tyrosine as an ADPr‐acceptor amino acid in RPS3A (Y155) and HPF1 (Y238) and demonstrated that trans‐modification of RPS3A is dependent on HPF1. We provide an ADPr‐site Localization Spectra Database (ADPr‐LSD), which contains 288 high‐quality ADPr‐modified peptide spectra, to serve as ADPr spectral references for correct ADPr‐site localizations. |
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| Keywords: | ADP‐ribosylation ARTD1/PARP1 genotoxic stress HPF1 tyrosine ADP‐ribosylation |
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