Proteomic investigations reveal a role for RNA processing factor THRAP3 in the DNA damage response

TY - JOUR

T1 - Proteomic investigations reveal a role for RNA processing factor THRAP3 in the DNA damage response

AU - Beli, Petra

AU - Lukashchuk, Natalia

AU - Wagner, Sebastian A

AU - Weinert, Brian T

AU - Olsen, Jesper V

AU - Baskcomb, Linda

AU - Mann, Matthias

AU - Jackson, Stephen P

AU - Choudhary, Chuna Ram

N1 - Copyright © 2012 Elsevier Inc. All rights reserved.

PY - 2012/4/27

Y1 - 2012/4/27

N2 - The regulatory networks of the DNA damage response (DDR) encompass many proteins and posttranslational modifications. Here, we use mass spectrometry-based proteomics to analyze the systems-wide response to DNA damage by parallel quantification of the DDR-regulated phosphoproteome, acetylome, and proteome. We show that phosphorylation-dependent signaling networks are regulated more strongly compared to acetylation. Among the phosphorylated proteins identified are many putative substrates of DNA-PK, ATM, and ATR kinases, but a majority of phosphorylated proteins do not share the ATM/ATR/DNA-PK target consensus motif, suggesting an important role of downstream kinases in amplifying DDR signals. We show that the splicing-regulator phosphatase PPM1G is recruited to sites of DNA damage, while the splicing-associated protein THRAP3 is excluded from these regions. Moreover, THRAP3 depletion causes cellular hypersensitivity to DNA-damaging agents. Collectively, these data broaden our knowledge of DNA damage signaling networks and highlight an important link between RNA metabolism and DNA repair.

AB - The regulatory networks of the DNA damage response (DDR) encompass many proteins and posttranslational modifications. Here, we use mass spectrometry-based proteomics to analyze the systems-wide response to DNA damage by parallel quantification of the DDR-regulated phosphoproteome, acetylome, and proteome. We show that phosphorylation-dependent signaling networks are regulated more strongly compared to acetylation. Among the phosphorylated proteins identified are many putative substrates of DNA-PK, ATM, and ATR kinases, but a majority of phosphorylated proteins do not share the ATM/ATR/DNA-PK target consensus motif, suggesting an important role of downstream kinases in amplifying DDR signals. We show that the splicing-regulator phosphatase PPM1G is recruited to sites of DNA damage, while the splicing-associated protein THRAP3 is excluded from these regions. Moreover, THRAP3 depletion causes cellular hypersensitivity to DNA-damaging agents. Collectively, these data broaden our knowledge of DNA damage signaling networks and highlight an important link between RNA metabolism and DNA repair.

KW - Cell Line, Tumor

KW - DNA Damage

KW - DNA Repair

KW - DNA-Binding Proteins

KW - HeLa Cells

KW - Humans

KW - Phosphoprotein Phosphatases

KW - Phosphorylation

KW - Proteomics

KW - Signal Transduction

KW - Transcription Factors

KW - Tumor Necrosis Factor-alpha

U2 - 10.1016/j.molcel.2012.01.026

DO - 10.1016/j.molcel.2012.01.026

M3 - Journal article

C2 - 22424773

SN - 1097-2765

VL - 46

SP - 212

EP - 225

JO - Molecular Cell

JF - Molecular Cell

IS - 2

ER -