Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

GEF-H1 upon microtubule destabilization drives ZBP1-dependent necroptosis

Cell Death & Differentiation (2026) Cite this article

Subjects

Abstract

It remains unclear for the role of microtubule-associated proteins or signaling in necroptosis. Here, we conducted high-throughput screening using a cytoskeleton compound library and identified that release of guanine nucleotide exchange factor-H1 (GEF-H1) upon microtubule destabilization triggers necroptosis. Immunoprecipitation and mass spectrometry revealed that GEF-H1 interacts with DExH-Box helicase 9 (DHX9) and protein-protein interaction network analysis indicated DHX9 is associated with necroptosis. Deficiency of either GEF-H1 or DHX9 significantly inhibits necroptosis. Mechanistically, upon microtubule destabilization, GEF-H1 is translocated to the nucleus and recruits DHX9 and RNA Pol II to the Z-DNA binding protein 1 (ZBP1) promoter region. Subsequently, increasing expression of ZBP1 drives necroptosis. In addition, GEF-H1 signaling upon microtubule destabilization also plays a positive role in lipopolysaccharide-induced cell death and inflammation. Collectively, these findings uncover an important role for GEF-H1 signaling in necroptosis and inflammation.

The alternative text for this image may have been generated using AI.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Microtubule destabilization triggers necroptosis.
The alternative text for this image may have been generated using AI.
Fig. 2: Inhibition of GEF-H1 restrains microtubule destabilization-mediated cell death.
The alternative text for this image may have been generated using AI.
Fig. 3: Knockout of GEF-H1 prominently inhibits necroptosis.
The alternative text for this image may have been generated using AI.
Fig. 4: Crosstalk between GEF-H1 and DHX9 triggers necroptosis.
The alternative text for this image may have been generated using AI.
Fig. 5: GEF-H1 interacts with DHX9 to synergistically upregulate ZBP1 expression.
The alternative text for this image may have been generated using AI.
Fig. 6: PH and Zinc-finger domain are required for GEF-H1-mediated ZBP1 expression.
The alternative text for this image may have been generated using AI.
Fig. 7: LPS activates GEF-H1 signaling and induces inflammation in vitro.
The alternative text for this image may have been generated using AI.
Fig. 8: GEF-H1 deficiency alleviates tissue damage and lethality induced by LPS and hvKP.
The alternative text for this image may have been generated using AI.

Similar content being viewed by others

Data availability

Data supporting the findings of this study are available from the corresponding author upon reasonable request. Original uncropped western blots are available as Supplementary Material.

References

  1. Mostowy S, Shenoy AR. The cytoskeleton in cell-autonomous immunity: structural determinants of host defence. Nat Rev Immunol. 2015;15:559–73.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  2. Joo E, Olson MF. Regulation and functions of the RhoA regulatory guanine nucleotide exchange factor GEF-H1. Small GTPases. 2021;12:358–71.

    Article  PubMed  CAS  Google Scholar 

  3. Kashyap AS, Fernandez-Rodriguez L, Zhao Y, Monaco G, Trefny MP, Yoshida N, et al. GEF-H1 signaling upon microtubule destabilization is required for dendritic cell activation and specific anti-tumor responses. Cell Rep. 2019;28:3367–80 e8.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  4. Chiang HS, Zhao Y, Song JH, Liu S, Wang N, Terhorst C, et al. GEF-H1 controls microtubule-dependent sensing of nucleic acids for antiviral host defenses. Nat Immunol. 2014;15:63–71.

    Article  PubMed  CAS  Google Scholar 

  5. Zhang X, Wu J, Liu Q, Li X, Li S, Chen J, et al. mtDNA-STING pathway promotes necroptosis-dependent enterocyte injury in intestinal ischemia reperfusion. Cell Death Dis. 2020;11:1050.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  6. Rucker AJ, Park CS, Li QJ, Moseman EA, Chan FK. Necroptosis stimulates interferon-mediated protective anti-tumor immunity. Cell Death Dis. 2024;15:403.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  7. Zhang X, Wu J, Liu Q, Li X, Yang Y, Wu L, et al. RIPK3-MLKL necroptotic signalling amplifies STING pathway and exacerbates lethal sepsis. Clin Transl Med. 2023;13:e1334.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  8. Basavaraju S, Mishra S, Jindal R, Kesavardhana S. Emerging role of ZBP1 in Z-RNA sensing, influenza virus-induced cell death, and pulmonary inflammation. mBio. 2022;13:e0040122.

    Article  PubMed  PubMed Central  Google Scholar 

  9. Galluzzi L, Kepp O, Chan FK, Kroemer G. Necroptosis: mechanisms and relevance to disease. Annu Rev Pathol. 2017;12:103–30.

    Article  PubMed  CAS  Google Scholar 

  10. Xie F, Wu D, Huang J, Liu X, Shen Y, Huang J, et al. ZBP1 condensate formation synergizes Z-NAs recognition and signal transduction. Cell Death Dis. 2024;15:487.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Malhotra J, Chiappori A, Fujioka N, Hanna NH, Feldman LE, Patel M, et al. Phase I/II trial of plinabulin in combination with nivolumab and ipilimumab in patients with recurrent small cell lung cancer (SCLC): Big ten cancer research consortium (BTCRC-LUN17-127) study. Lung Cancer. 2024;195:107932.

    Article  PubMed  CAS  Google Scholar 

  12. Birkenfeld J, Nalbant P, Yoon SH, Bokoch GM. Cellular functions of GEF-H1, a microtubule-regulated Rho-GEF: is altered GEF-H1 activity a crucial determinant of disease pathogenesis? Trends Cell Biol. 2008;18:210–9.

    Article  PubMed  CAS  Google Scholar 

  13. Kuriakose T, Kanneganti TD. ZBP1: innate sensor regulating cell death and inflammation. Trends Immunol. 2018;39:123–34.

    Article  PubMed  CAS  Google Scholar 

  14. Imai T, Lin J, Kaya GG, Ju E, Kondylis V, Kelepouras K, et al. The RIPK1 death domain restrains ZBP1- and TRIF-mediated cell death and inflammation. Immunity. 2024;57:1497–513.e6.

    Article  PubMed  CAS  Google Scholar 

  15. Lai K, Wang J, Lin S, Chen Z, Lin G, Ye K, et al. Sensing of mitochondrial DNA by ZBP1 promotes RIPK3-mediated necroptosis and ferroptosis in response to diquat poisoning. Cell Death Differ. 2024;31:635–50.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  16. Koerner L, Wachsmuth L, Kumari S, Schwarzer R, Wagner T, Jiao H, et al. ZBP1 causes inflammation by inducing RIPK3-mediated necroptosis and RIPK1 kinase activity-independent apoptosis. Cell Death Differ. 2024;31:938–53.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  17. Ren X, Wang D, Zhang G, Zhou T, Wei Z, Yang Y, et al. Nucleic DHX9 cooperates with STAT1 to transcribe interferon-stimulated genes. Sci Adv. 2023;9:eadd5005.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  18. Nakajima T, Uchida C, Anderson SF, Lee CG, Hurwitz J, Parvin JD, et al. RNA helicase A mediates association of CBP with RNA polymerase II. Cell. 1997;90:1107–12.

    Article  PubMed  CAS  Google Scholar 

  19. Lu J, Wu T, Zhang B, Liu S, Song W, Qiao J, et al. Types of nuclear localization signals and mechanisms of protein import into the nucleus. Cell Commun Signal. 2021;19:60.

  20. Zhao Y, Zagani R, Park SM, Yoshida N, Shah P, Reinecker HC. Microbial recognition by GEF-H1 controls IKKepsilon mediated activation of IRF5. Nat Commun. 2019;10:1349.

    Article  PubMed  PubMed Central  Google Scholar 

  21. Ren X, Liu Q, Zhou P, Zhou T, Wang D, Mei Q, et al. DHX9 maintains epithelial homeostasis by restraining R-loop-mediated genomic instability in intestinal stem cells. Nat Commun. 2024;15:3080.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. Delgado M, Rainwater RR, Heflin B, Urbaniak A, Butler K, Davidson M, et al. Primary acute lymphoblastic leukemia cells are susceptible to microtubule depolymerization in G1 and M phases through distinct cell death pathways. J Biol Chem. 2022;298:101939.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Hugle M, Belz K, Fulda S. Identification of synthetic lethality of PLK1 inhibition and microtubule-destabilizing drugs. Cell Death Differ. 2015;22:1946–56.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  24. Sebastian J, Rathinasamy K. Microtubules and cell division: potential pharmacological targets in cancer therapy. Curr Drug Targets. 2023;24:889–918.

    Article  PubMed  CAS  Google Scholar 

  25. Jiao A, Sun C, Wang X, Lei L, Liu H, Li W, et al. DExD/H-box helicase 9 intrinsically controls CD8(+) T cell-mediated antiviral response through noncanonical mechanisms. Sci Adv. 2022;8:eabk2691.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  26. Fuller-Pace FV. DExD/H box RNA helicases: multifunctional proteins with important roles in transcriptional regulation. Nucleic Acids Res. 2006;34:4206–15.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  27. Yang BZ, Liu MY, Chiu KL, Chien YL, Cheng CA, Chen YL, et al. DHX9 SUMOylation is required for the suppression of R-loop-associated genome instability. Nat Commun. 2024;15:6009.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  28. Castro J, Daniels MH, Brennan D, Johnston B, Gotur D, Lee YT, et al. A potent, selective, small-molecule inhibitor of DHX9 abrogates proliferation of microsatellite instable cancers with deficient mismatch repair. Cancer Res. 2024;85:758–76.

  29. Ng YC, Chung WC, Kang HR, Cho HJ, Park EB, Kang SJ, et al. A DNA-sensing-independent role of a nuclear RNA helicase, DHX9, in stimulation of NF-kappaB-mediated innate immunity against DNA virus infection. Nucleic Acids Res. 2018;46:9011–26.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  30. Zhu S, Ding S, Wang P, Wei Z, Pan W, Palm NW, et al. Nlrp9b inflammasome restricts rotavirus infection in intestinal epithelial cells. Nature. 2017;546:667–70.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  31. Singer M, Deutschman CS, Seymour CW, Shankar-Hari M, Annane D, Bauer M, et al. The third international consensus definitions for sepsis and septic shock (sepsis-3). JAMA. 2016;315:801–10.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  32. Wu J, Liu Q, Zhang X, Tan M, Li X, Liu P, et al. The interaction between STING and NCOA4 exacerbates lethal sepsis by orchestrating ferroptosis and inflammatory responses in macrophages. Cell Death Dis. 2022;13:653.

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  33. Wu J, Liu Q, Zhang X, Wu X, Zhao Y, Ren J. STING-dependent induction of lipid peroxidation mediates intestinal ischemia-reperfusion injury. Free Radical Biol Med. 2021;163:135–40.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We are grateful to Prof. Jiahuai Han (Xiamen University) for providing experimental materials.

Funding

National Natural Science Foundation of China (82272237, 82430083, 82072149), Basic Research Program of Jiangsu (BK20250108) and Natural Science Foundation of Jiangsu Province (CXZX202217).

Author information

Author notes

  1. These authors contributed equally: Xufei Zhang, Xuanheng Li, Yiyu Yang.

Authors and Affiliations

  1. Department of General Surgery, Nanjing Drum Tower Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China

    Xufei Zhang

  2. Research Institute of General Surgery, Jinling Hospital, School of Medicine, Southeast University, Nanjing, China

    Xufei Zhang, Yiyu Yang, Liting Deng, Jianan Ren & Xiuwen Wu

  3. Research Institute of General Surgery, Jinling Hospital, Affiliated Hospital of Medical School, Nanjing University, Nanjing, China

    Xuanheng Li, Jinjian Huang, Jianan Ren & Xiuwen Wu

  4. Department of Critical Care Medicine, Affiliated Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China

    Jie Wu

  5. Department of General Surgery, Affiliated Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China

    Qinjie Liu

  6. Research Center of Surgery, BenQ Medical Center, the Affiliated BenQ Hospital of Nanjing Medical University, Nanjing, China

    Yun Zhao

Authors
  1. Xufei Zhang
    View author publications

    Search author on:PubMed Google Scholar

  2. Xuanheng Li
    View author publications

    Search author on:PubMed Google Scholar

  3. Yiyu Yang
    View author publications

    Search author on:PubMed Google Scholar

  4. Jie Wu
    View author publications

    Search author on:PubMed Google Scholar

  5. Qinjie Liu
    View author publications

    Search author on:PubMed Google Scholar

  6. Liting Deng
    View author publications

    Search author on:PubMed Google Scholar

  7. Jinjian Huang
    View author publications

    Search author on:PubMed Google Scholar

  8. Yun Zhao
    View author publications

    Search author on:PubMed Google Scholar

  9. Jianan Ren
    View author publications

    Search author on:PubMed Google Scholar

  10. Xiuwen Wu
    View author publications

    Search author on:PubMed Google Scholar

Contributions

XW Wu, JA Ren, Y Zhao, and XF Zhang designed this research; XF Zhang, XH Li, YY Yang, J Wu, and QJ Liu conducted the research; XF Zhang, LT Deng, and JJ Huang performed the statistical analysis; XF Zhang, XW Wu, JA Ren, and Y Zhao wrote and revised the paper.

Corresponding authors

Correspondence to Yun Zhao, Jianan Ren or Xiuwen Wu.

Ethics declarations

Competing interests

The authors declare no competing interests.

Ethics approval and consent to participate

All procedures for animal experiments were approved by Jinling Hospital Animal Care Committee. The collection of human intestinal tissue was approved by the Institutional Review Board Ethics Committee at Jinling Hospital. Before collecting samples, informed consent was obtained from all human participants. All methods were performed in accordance with the relevant guidelines and regulations.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Zhang, X., Li, X., Yang, Y. et al. GEF-H1 upon microtubule destabilization drives ZBP1-dependent necroptosis. Cell Death Differ (2026). https://doi.org/10.1038/s41418-026-01711-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Version of record:

  • DOI: https://doi.org/10.1038/s41418-026-01711-y

Search

Advanced search

Quick links