Combined experimental and computational analysis of DNA damage signaling reveals context‐dependent roles for Erk in apoptosis and G1/S arrest after genotoxic stress

Molecular Systems Biology

Andrea R Tentner¹  Michael J Lee¹  Gerry J Ostheimer¹  Leona D Samson¹  Douglas A Lauffenburger¹ 

[1] Departments of Biology and Biological Engineering, David H Koch Institute for Integrative Cancer Research at MIT, Massachusetts Institute of Technology, Cambridge, MA, USA

关键词: apoptosis;    cell‐cycle checkpoint;    DNA damage;    Erk;    signal transduction;   

DOI  :  10.1038/msb.2012.1

来源: Wiley

【 摘 要 】

Abstract

Following DNA damage, cells display complex multi-pathway signaling dynamics that connect cell-cycle arrest and DNA repair in G1, S, or G2/M phase with phenotypic fate decisions made between survival, cell-cycle re-entry and proliferation, permanent cell-cycle arrest, or cell death. How these phenotypic fate decisions are determined remains poorly understood, but must derive from integrating genotoxic stress signals together with inputs from the local microenvironment. To investigate this in a systematic manner, we undertook a quantitative time-resolved cell signaling and phenotypic response study in U2OS cells receiving doxorubicin-induced DNA damage in the presence or absence of TNFα co-treatment; we measured key nodes in a broad set of DNA damage signal transduction pathways along with apoptotic death and cell-cycle regulatory responses. Two relational modeling approaches were then used to identify network-level relationships between signals and cell phenotypic events: a partial least squares regression approach and a complementary new technique which we term ‘time-interval stepwise regression.’ Taken together, the results from these analysis methods revealed complex, cytokine-modulated inter-relationships among multiple signaling pathways following DNA damage, and identified an unexpected context-dependent role for Erk in both G1/S arrest and apoptotic cell death following treatment with this commonly used clinical chemotherapeutic drug.

Synopsis

Data-driven modeling was used to analyze the complex signaling dynamics that connect DNA repair with cell survival, cell-cycle arrest, or apoptosis. This analysis revealed an unexpected role for Erk in G1/S arrest and apoptotic cell death following

• The cell-fate choice between apoptosis and cell-cycle arrest after doxorubicin treatment is dose dependent and modulated by TNF.
• An early, transient G1/S arrest following doxorubicin treatment acts as a cell-fate decision point between re-entry into S-phase and apoptotic cell death.
• Two complementary relational modeling methods identify the proliferation and survival signal, ERK, as potentially causal for the G1/S arrest and death phenotypes.
• Inhibitor experiments validate a role for ERK in maintenance of the early, transient arrest and in promotion of apoptosis from this arrested state following doxorubicin-induced DNA damage.

【 授权许可】

CC BY-NC-SA   
Copyright © 2012 EMBO and Macmillan Publishers Limited

Creative Commons Attribution License, which permits distribution, and reproduction in any medium, provided the original author and source are credited. This license does not permit commercial exploitation without specific permission.