Uniform Widespread Nuclear Phosphorylation of Histone H2AX Is an Indicator of Lethal DNA Replication Stress

Cancers

Audrey Stoessel¹  Marc Vigneron¹  Eric Moeglin¹  Etienne Weiss¹  Manuela Chiper¹  Dominique Desplancq¹  Laszlo Tora²  Sascha Conic²  Mustapha Oulad-Abdelghani²  Pascal Didier³ 

[1]Biotechnologie et Signalisation Cellulaire, UMR 7242, CNRS/Université de Strasbourg, Boulevard S. Brant, 67412 Illlkirch, France

[2]Institut de Génétique et de Biologie Moléculaire et Cellulaire, 67404 Illkirch, France

[3]Laboratoire de Bioimagerie et Pathologies, UMR 7213, CNRS/Université de Strasbourg, Route du Rhin, 67401 Illkirch, France

关键词: histone variant;    H2AX phosphorylation;    γ-H2AX;    pan-nuclear pattern;    monoclonal antibody;    cancer cells;    cell death;    replication stress;    chemotherapy;    H2AFX gene;    knock-out;   

DOI  :  10.3390/cancers11030355

来源: DOAJ

【 摘 要 】

Phosphorylated histone H2AX (γ-H2AX), a central player in the DNA damage response (DDR), serves as a biomarker of DNA double-strand break repair. Although DNA damage is generally visualized by the formation of γ-H2AX foci in injured nuclei, it is unclear whether the widespread uniform nuclear γ-H2AX (called pan-nuclear) pattern occurring upon intense replication stress (RS) is linked to DDR. Using a novel monoclonal antibody that binds exclusively to the phosphorylated C-terminus of H2AX, we demonstrate that H2AX phosphorylation is systematically pan-nuclear in cancer cells stressed with RS-inducing drugs just before they die. The pan-nuclear γ-H2AX pattern is abolished by inhibition of the DNA-PK kinase. Cell death induction of cancer cells treated with increasing combinations of replication and kinase (ATR and Chk1) inhibitory drugs was proportional to the appearance of pan-nuclear γ-H2AX pattern. Delivery of labeled anti-γ-H2AX Fabs in stressed cells demonstrated at a single cell level that pan-nuclear γ-H2AX formation precedes irreversible cell death. Moreover, we show that H2AX is not required for RS-induced cell death in HeLa cells. Thus, the nuclear-wide formation of γ-H2AX is an incident of RS-induced cell death and, thus, the pan nuclear H2AX pattern should be regarded as an indicator of lethal RS-inducing drug efficacy.

【 授权许可】

Unknown