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Comprehensive ADP-ribosylome analysis identifies tyrosine as an ADP-ribose acceptor site

ABSTRACT: Despite recent mass spectrometry (MS)-based breakthroughs, comprehensive ADP-ribose (ADPr)-site identification and 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. 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.

INSTRUMENT(S): Orbitrap Fusion, Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Permanent Cell Line Cell

SUBMITTER: Michael Hottiger

LAB HEAD: Michael O. Hottiger

PROVIDER: PXD009948 | Pride | 2018-06-28

REPOSITORIES: Pride

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Comprehensive ADP-ribosylome analysis identifies tyrosine as an ADP-ribose acceptor site.

Leslie Pedrioli Deena M DM Leutert Mario M Bilan Vera V Nowak Kathrin K Gunasekera Kapila K Ferrari Elena E Imhof Ralph R Malmström Lars L Hottiger Michael O MO

EMBO reports 20180628 8

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 <i>in vivo</i> ADPr-acceptor amino acid. MS analyses of <i>in vitro</i> ADP-ribosylated proteins confirmed tyrosine as an ADPr-acceptor amino acid in RPS ...[more]

PMID: 29954836

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