PortalPublikationenQuantitative proteomics unveiled: Regulation of DNA double s...

Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1

Standard

Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1. / Myllynen, Laura; Kwiatkowski, Marcel; Gleißner, Lisa; Riepen, Britta; Hoffer, Konstantin; Wurlitzer, Marcus; Petersen, Cordula; Dikomey, Ekkehard; Rothkamm, Kai; Schlüter, Hartmut; Kriegs, Malte.

in: RADIOTHER ONCOL, Jahrgang 116, Nr. 3, 09.2015, S. 423-30.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Myllynen, L, Kwiatkowski, M, Gleißner, L, Riepen, B, Hoffer, K, Wurlitzer, M, Petersen, C, Dikomey, E, Rothkamm, K, Schlüter, H & Kriegs, M 2015, 'Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1', RADIOTHER ONCOL, Jg. 116, Nr. 3, S. 423-30. https://doi.org/10.1016/j.radonc.2015.09.018

APA

Myllynen, L., Kwiatkowski, M., Gleißner, L., Riepen, B., Hoffer, K., Wurlitzer, M., Petersen, C., Dikomey, E., Rothkamm, K., Schlüter, H., & Kriegs, M. (2015). Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1. RADIOTHER ONCOL, 116(3), 423-30. https://doi.org/10.1016/j.radonc.2015.09.018

Vancouver

Myllynen L, Kwiatkowski M, Gleißner L, Riepen B, Hoffer K, Wurlitzer M et al. Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1. RADIOTHER ONCOL. 2015 Sep;116(3):423-30. https://doi.org/10.1016/j.radonc.2015.09.018

Bibtex

@article{e894133493d34d2cbc652ef2d4db7c84,
title = "Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1",
abstract = "BACKGROUND: EGFR inhibition blocks DNA double strand break (DSB) repair but the detailed mechanisms are still unclear. We asked whether EGFR inhibition blocks DSB repair by reducing the X-ray-induced phosphorylation of repair proteins using a phosphoproteomic approach.MATERIALS AND METHODS: Using UT-SCC5 and SAS head and neck cancer cells we established a differential phosphoproteomic approach for quantitative analysis of DNA repair proteins by stable isotope labeling with amino acids. Nuclear phosphoproteins were isolated and analyzed by liquid chromatography/tandem mass spectrometry. Erlotinib, PD98059 and olaparib were used to inhibit EGFR, MEK1/2 and PARP1, respectively. PARP1 was knocked down by siRNA. DSB repair was measured by quantifying residual 53BP1 foci.RESULTS: Over 150 nuclear phosphoproteins were quantified after irradiation, including 24 DNA repair proteins. Two of these, including PARP1, were consistently reduced in both cell lines upon erlotinib treatment. PARP1 inhibition or knock-down and EGFR inhibition resulted in an analog number of residual foci which was not further increased by combination of both strategies. MEK1/2 inhibition with or without blockage of EGFR or PARP1 caused similar effects.CONCLUSION: We have established a powerful, quantitative phosphoproteomic approach to investigate regulatory mechanisms in DSB repair, dependent on protein phosphorylation after irradiation. Using this approach we have identified PARP1 as a mediator of EGFR/MEK-dependent regulation of DSB repair.",
author = "Laura Myllynen and Marcel Kwiatkowski and Lisa Glei{\ss}ner and Britta Riepen and Konstantin Hoffer and Marcus Wurlitzer and Cordula Petersen and Ekkehard Dikomey and Kai Rothkamm and Hartmut Schl{\"u}ter and Malte Kriegs",
note = "Copyright {\textcopyright} 2015 Elsevier Ireland Ltd. All rights reserved.",
year = "2015",
month = sep,
doi = "10.1016/j.radonc.2015.09.018",
language = "English",
volume = "116",
pages = "423--30",
journal = "RADIOTHER ONCOL",
issn = "0167-8140",
publisher = "Elsevier Ireland Ltd",
number = "3",

}

RIS

TY - JOUR

T1 - Quantitative proteomics unveiled: Regulation of DNA double strand break repair by EGFR involves PARP1

AU - Myllynen, Laura

AU - Kwiatkowski, Marcel

AU - Gleißner, Lisa

AU - Riepen, Britta

AU - Hoffer, Konstantin

AU - Wurlitzer, Marcus

AU - Petersen, Cordula

AU - Dikomey, Ekkehard

AU - Rothkamm, Kai

AU - Schlüter, Hartmut

AU - Kriegs, Malte

N1 - Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

PY - 2015/9

Y1 - 2015/9

N2 - BACKGROUND: EGFR inhibition blocks DNA double strand break (DSB) repair but the detailed mechanisms are still unclear. We asked whether EGFR inhibition blocks DSB repair by reducing the X-ray-induced phosphorylation of repair proteins using a phosphoproteomic approach.MATERIALS AND METHODS: Using UT-SCC5 and SAS head and neck cancer cells we established a differential phosphoproteomic approach for quantitative analysis of DNA repair proteins by stable isotope labeling with amino acids. Nuclear phosphoproteins were isolated and analyzed by liquid chromatography/tandem mass spectrometry. Erlotinib, PD98059 and olaparib were used to inhibit EGFR, MEK1/2 and PARP1, respectively. PARP1 was knocked down by siRNA. DSB repair was measured by quantifying residual 53BP1 foci.RESULTS: Over 150 nuclear phosphoproteins were quantified after irradiation, including 24 DNA repair proteins. Two of these, including PARP1, were consistently reduced in both cell lines upon erlotinib treatment. PARP1 inhibition or knock-down and EGFR inhibition resulted in an analog number of residual foci which was not further increased by combination of both strategies. MEK1/2 inhibition with or without blockage of EGFR or PARP1 caused similar effects.CONCLUSION: We have established a powerful, quantitative phosphoproteomic approach to investigate regulatory mechanisms in DSB repair, dependent on protein phosphorylation after irradiation. Using this approach we have identified PARP1 as a mediator of EGFR/MEK-dependent regulation of DSB repair.

AB - BACKGROUND: EGFR inhibition blocks DNA double strand break (DSB) repair but the detailed mechanisms are still unclear. We asked whether EGFR inhibition blocks DSB repair by reducing the X-ray-induced phosphorylation of repair proteins using a phosphoproteomic approach.MATERIALS AND METHODS: Using UT-SCC5 and SAS head and neck cancer cells we established a differential phosphoproteomic approach for quantitative analysis of DNA repair proteins by stable isotope labeling with amino acids. Nuclear phosphoproteins were isolated and analyzed by liquid chromatography/tandem mass spectrometry. Erlotinib, PD98059 and olaparib were used to inhibit EGFR, MEK1/2 and PARP1, respectively. PARP1 was knocked down by siRNA. DSB repair was measured by quantifying residual 53BP1 foci.RESULTS: Over 150 nuclear phosphoproteins were quantified after irradiation, including 24 DNA repair proteins. Two of these, including PARP1, were consistently reduced in both cell lines upon erlotinib treatment. PARP1 inhibition or knock-down and EGFR inhibition resulted in an analog number of residual foci which was not further increased by combination of both strategies. MEK1/2 inhibition with or without blockage of EGFR or PARP1 caused similar effects.CONCLUSION: We have established a powerful, quantitative phosphoproteomic approach to investigate regulatory mechanisms in DSB repair, dependent on protein phosphorylation after irradiation. Using this approach we have identified PARP1 as a mediator of EGFR/MEK-dependent regulation of DSB repair.

U2 - 10.1016/j.radonc.2015.09.018

DO - 10.1016/j.radonc.2015.09.018

M3 - SCORING: Journal article

C2 - 26422459

VL - 116

SP - 423

EP - 430

JO - RADIOTHER ONCOL

JF - RADIOTHER ONCOL

SN - 0167-8140

IS - 3

ER -