Temporal and site-specific adp-ribosylation dynamics upon different genotoxic stresses
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Temporal and site-specific adp-ribosylation dynamics upon different genotoxic stresses. / Buch-Larsen, Sara C.; Rebak, Alexandra K.L.F.S.; Hendriks, Ivo A.; Nielsen, Michael L.
I: Cells, Bind 10, Nr. 11, 2927, 2021.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Temporal and site-specific adp-ribosylation dynamics upon different genotoxic stresses
AU - Buch-Larsen, Sara C.
AU - Rebak, Alexandra K.L.F.S.
AU - Hendriks, Ivo A.
AU - Nielsen, Michael L.
N1 - Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021
Y1 - 2021
N2 - The DNA damage response revolves around transmission of information via post-translational modifications, including reversible protein ADP-ribosylation. Here, we applied a massspectrometry-based Af1521 enrichment technology for the identification and quantification of ADPribosylation sites as a function of various DNA damage stimuli and time. In total, we detected 1681 ADP-ribosylation sites residing on 716 proteins in U2OS cells and determined their temporal dynamics after exposure to the genotoxins H2O2 and MMS. Intriguingly, we observed a widespread but low-abundance serine ADP-ribosylation response at the earliest time point, with later time points centered on increased modification of the same sites. This suggests that early serine ADPribosylation events may serve as a platform for an integrated signal response. While treatment with H2O2 and MMS induced homogenous ADP-ribosylation responses, we observed temporal differences in the ADP-ribosylation site abundances. Exposure to MMS-induced alkylating stress induced the strongest ADP-ribosylome response after 30 min, prominently modifying proteins involved in RNA processing, whereas in response to H2O2-induced oxidative stress ADP-ribosylation peaked after 60 min, mainly modifying proteins involved in DNA damage pathways. Collectively, the dynamic ADP-ribosylome presented here provides a valuable insight into the temporal cellular regulation of ADP-ribosylation in response to DNA damage.
AB - The DNA damage response revolves around transmission of information via post-translational modifications, including reversible protein ADP-ribosylation. Here, we applied a massspectrometry-based Af1521 enrichment technology for the identification and quantification of ADPribosylation sites as a function of various DNA damage stimuli and time. In total, we detected 1681 ADP-ribosylation sites residing on 716 proteins in U2OS cells and determined their temporal dynamics after exposure to the genotoxins H2O2 and MMS. Intriguingly, we observed a widespread but low-abundance serine ADP-ribosylation response at the earliest time point, with later time points centered on increased modification of the same sites. This suggests that early serine ADPribosylation events may serve as a platform for an integrated signal response. While treatment with H2O2 and MMS induced homogenous ADP-ribosylation responses, we observed temporal differences in the ADP-ribosylation site abundances. Exposure to MMS-induced alkylating stress induced the strongest ADP-ribosylome response after 30 min, prominently modifying proteins involved in RNA processing, whereas in response to H2O2-induced oxidative stress ADP-ribosylation peaked after 60 min, mainly modifying proteins involved in DNA damage pathways. Collectively, the dynamic ADP-ribosylome presented here provides a valuable insight into the temporal cellular regulation of ADP-ribosylation in response to DNA damage.
KW - ADP-ribosylation
KW - Af1521
KW - DNA damage
KW - Mass spectrometry
KW - PARP
KW - Post-translational modification
KW - Proteomics
U2 - 10.3390/cells10112927
DO - 10.3390/cells10112927
M3 - Journal article
C2 - 34831150
AN - SCOPUS:85117947962
VL - 10
JO - Cells
JF - Cells
SN - 2073-4409
IS - 11
M1 - 2927
ER -
ID: 284289311