Actin networks modulate heterogenous NF-κB dynamics in response to TNFα

  1. Francesca Butera  Is a corresponding author
  2. Julia E Sero
  3. Lucas G Dent  Is a corresponding author
  4. Chris Bakal  Is a corresponding author
  1. Institute of Cancer Research, United Kingdom
  2. University of Bath, United Kingdom

Abstract

The canonical NF-κB transcription factor RELA is a master regulator of immune and stress responses and is upregulated in PDAC tumours. In this study, we characterised previously unexplored endogenous RELA-GFP dynamics in PDAC cell lines through live single cell imaging. Our observations revealed that TNFα stimulation induces rapid, sustained, and non-oscillatory nuclear translocation of RELA. Through Bayesian analysis of single cell datasets with variation in nuclear RELA, we predicted that RELA heterogeneity in PDAC cell lines is dependent on F-actin dynamics. RNA-seq analysis identified distinct clusters of RELA-regulated gene expression in PDAC cells, including TNFα-induced RELA upregulation of the actin regulators NUAK2 and ARHGAP31. Further, siRNA-mediated depletion of ARHGAP31 and NUAK2 altered TNFα-stimulated nuclear RELA dynamics in PDAC cells, establishing a novel negative feedback loop that regulates RELA activation by TNFα. Additionally, we characterised the NF-κB pathway in PDAC cells, identifying how NF-κB/IκB proteins genetically and physically interact with RELA in the absence or presence of TNFα. Taken together, we provide computational and experimental support for interdependence between the F-actin network and the NF-κB pathway with RELA translocation dynamics in PDAC.

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All data generated for this study have been included as source data files.

Article and author information

Author details

  1. Francesca Butera

    Division of Cancer Biology, Institute of Cancer Research, London, United Kingdom
    For correspondence
    frankie.butera@mcri.edu.au
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-6606-4678
  2. Julia E Sero

    Biology and Biochemistry Department, University of Bath, Bath, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  3. Lucas G Dent

    Division of Cancer Biology, Institute of Cancer Research, London, United Kingdom
    For correspondence
    lucas.dent@icr.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8573-4617
  4. Chris Bakal

    Division of Cancer Biology, Institute of Cancer Research, London, United Kingdom
    For correspondence
    cbakal@icr.ac.uk
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0413-6744

Funding

Cancer Research UK (S_3567)

  • Francesca Butera

Cancer Research UK supported by Stand Up to Cancer UK (C37275)

  • Chris Bakal

Cancer Research UK supported by Stand Up to Cancer UK (A20146)

  • Chris Bakal

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2024, Butera et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Francesca Butera
  2. Julia E Sero
  3. Lucas G Dent
  4. Chris Bakal
(2024)
Actin networks modulate heterogenous NF-κB dynamics in response to TNFα
eLife 13:e86042.
https://doi.org/10.7554/eLife.86042

Share this article

https://doi.org/10.7554/eLife.86042

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