Abstract
ADP-ribosylation (ADPr) signaling plays a crucial role in DNA damage response. Inhibitors against the main enzyme catalyzing ADPr after DNA damage, poly(ADP-ribose) polymerase 1 (PARP1), are used to treat patients with breast cancer harboring BRCA1/2 mutations. However, resistance to PARP inhibitors (PARPi) is a major obstacle in treating patients. To understand the role of ADPr in PARPi sensitivity, we use liquid chromatography-tandem mass spectrometry (LC-MS/MS) to analyze ADPr in six breast cancer cell lines exhibiting different PARPi sensitivities. We identify 1,632 sites on 777 proteins across all cell lines, primarily on serine residues, with site-specific overlap of targeted residues across DNA-damage-related proteins across all cell lines, demonstrating high conservation of serine ADPr-signaling networks upon DNA damage. Furthermore, we observe site-specific differences in ADPr intensities in PARPi-sensitive BRCA mutants and unique ADPr sites in PARPi-resistant BRCA-mutant HCC1937 cells, which have low poly(ADP-ribose) glycohydrolase (PARG) levels and longer ADPr chains on PARP1.
| Originalsprog | Engelsk |
|---|---|
| Artikelnummer | 114433 |
| Tidsskrift | Cell Reports |
| Vol/bind | 43 |
| Udgave nummer | 7 |
| Antal sider | 23 |
| ISSN | 2211-1247 |
| DOI | |
| Status | Udgivet - 2024 |
Bibliografisk note
Funding Information:We thank the lab of Michael O. Hottiger for the expression and purification of recombinant human PARG (University of Zurich), and we thank members of the Novo Nordisk Foundation Center for Protein Research (NNF-CPR) Mass Spectrometry Platform, as well as Ivo Hendriks and Patrick Ruether for instrument support and technical assistance. We thank the lab of Jesper Olsen (NNF-CPR) for providing frozen vials of the breast cancer cell lines used in this study. We thank members of the M.L.N. lab for support. This study was supported by NNF-CPR, the Novo Nordisk Foundation (grant agreements NNF14CC0001 and NNF13OC0006477), the Danish Council for Independent Research (2032-00311B), and the Danish Cancer Institute (R325-A18824) to M.L.N. H.A.A. is supported by the Novo Nordisk Foundation Copenhagen Bioscience PhD program (grant agreement NNF19SA0035440). A.G.P. and Z.S. are supported by the Breast Cancer Research Foundation (BCRF-22-159), Kr\u00E6ftens Bek\u00E6mpelse (R325-A18809, R281-16566, and R342-A19788), Det Frie Forskningsr\u00E5d, and Sundhed og Sygdom (2034-00205B). Conceptualization, M.L.N, H.A.A. and S.C.B.-L.; investigation, H.A.A. H.C. and A.G.P.; formal analysis, H.A.A. M.M. and M.L.-P.; visualization, H.A.A. A.G.P. and M.L.-P.; writing \u2013 original draft, H.A.A. and A.G.P.; writing \u2013 review and editing, H.A.A. M.M. S.C.B.-L. and M.L.N.; supervision, M.L.N. M.L.-P. and Z.S.; funding acquisition, M.L.N. and Z.S. The authors declare no competing interests.
Funding Information:
We thank the lab of Michael O. Hottiger for the expression and purification of recombinant human PARG ( University of Zurich ), and we thank members of the Novo Nordisk Foundation Center for Protein Research (NNF-CPR) Mass Spectrometry Platform, as well as Ivo Hendriks and Patrick Ruether for instrument support and technical assistance. We thank the lab of Jesper Olsen (NNF-CPR) for providing frozen vials of the breast cancer cell lines used in this study. We thank members of the M.L.N. lab for support. This study was supported by NNF-CPR , the Novo Nordisk Foundation (grant agreements NNF14CC0001 and NNF13OC0006477 ), the Danish Council for Independent Research ( 2032-00311B ), and the Danish Cancer Institute ( R325-A18824 ) to M.L.N. H.A.A. is supported by the Novo Nordisk Foundation Copenhagen Bioscience PhD program (grant agreement NNF19SA0035440 ). A.G.P. and Z.S. are supported by the Breast Cancer Research Foundation ( BCRF-22-159 ), Kr\u00E6ftens Bek\u00E6mpelse ( R325-A18809 , R281-16566 , and R342-A19788 ), Det Frie Forskningsr\u00E5d, and Sundhed og Sygdom ( 2034-00205B ).
Publisher Copyright:
© 2024 The Author(s)