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ATR-mediated phosphorylation of RIPK1 inhibits DNA damage-induced necroptosis

Shen-Nan Shi1, Qiuyang Xu1, Xiaofei Jiao1

  • 1Department of Gynecological Oncology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China; National Clinical Research Center for Obstetrics and Gynecology, Cancer Biology Research Center (Key Laboratory of the Ministry of Education), Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.

Biochemical Pharmacology
|June 25, 2025
PubMed

Insights

Ataxia telangiectasia and rad3-related protein (ATR) suppresses DNA damage-induced necroptosis in ovarian cancer by inhibiting RIPK1. Targeting this interaction may enhance chemotherapy effectiveness and improve patient outcomes.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Cell Death Pathways

Background:

  • DNA damage during chemotherapy can trigger necroptosis, a programmed cell death pathway, offering a treatment strategy for epithelial ovarian cancer.
  • Ataxia telangiectasia and rad3-related protein (ATR) is a key kinase in DNA damage checkpoints, regulating repair and cell fate.
  • The precise mechanisms by which ATR influences necroptosis, particularly in the context of DNA damage, are not fully elucidated.

Purpose of the Study:

  • To investigate the role of ATR in regulating receptor-interacting serine/threonine-protein kinase 1 (RIPK1)-dependent necroptosis following DNA damage.
  • To elucidate the molecular mechanism through which ATR controls RIPK1 activation and necroptosis.
  • To determine the therapeutic implications of the ATR-RIPK1 interaction in ovarian cancer chemosensitivity.

Main Methods:

  • Investigated the interaction between ATR and RIPK1 using biochemical assays.
  • Utilized site-directed mutagenesis (S335A) to assess the role of RIPK1 phosphorylation by ATR.
  • Employed RIPK1 knockout and complementation systems in ovarian cancer cell lines to evaluate necroptosis and chemosensitivity.

Main Results:

  • ATR directly binds to RIPK1 and inhibits its activation and downstream necrosome formation by phosphorylating RIPK1 at Ser335 (S335).
  • The S335A mutation in RIPK1 relieved ATR-mediated inhibition, leading to enhanced necroptosis.
  • ATR-mediated phosphorylation of RIPK1 at S335 promoted chemoresistance in ovarian cancer cells, whereas the S335A mutation significantly increased chemosensitivity, cell death, and reduced viability.

Conclusions:

  • ATR-dependent phosphorylation of RIPK1 at S335 is a critical mechanism suppressing DNA damage-induced necroptosis and conferring chemoresistance in ovarian cancer.
  • Targeting the ATR-RIPK1 interaction, specifically by preventing S335 phosphorylation, represents a potential therapeutic strategy to enhance necroptosis and improve chemosensitivity in ovarian cancer treatment.
Keywords:
ATRChemosensitivityDNA damageNecroptosisOvarian cancerRIPK1

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