Actin networks modulate heterogeneous NF-[kappa]B dynamics in response to TNF[alpha]

The canonical NF-[kappa]B transcription factor RELA is a master regulator of immune and stress responses and is upregulated in pancreatic ductal adenocardinoma (PDAC) tumours. In this study, we characterised previously unexplored endogenous RELA-GFP dynamics in PDAC cell lines through live single-ce...

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Bibliographic Details
Published in:eLife 2024-08, Vol.13
Main Authors: Butera, Francesca, Sero, Julia E, Dent, Lucas G, Bakal, Chris
Format: Article
Language:English
Subjects:
Actin
Gene expression
Genes
Muscle proteins
Neomycin
RNA
Tumor necrosis factor
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Summary:The canonical NF-[kappa]B transcription factor RELA is a master regulator of immune and stress responses and is upregulated in pancreatic ductal adenocardinoma (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 TNF[alpha] 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 TNF[alpha]-induced RELA upregulation of the actin regulators NUAK2 and ARHGAP31. Further, siRNA-mediated depletion of ARHGAP31 and NUAK2 altered TNF[alpha]-stimulated nuclear RELA dynamics in PDAC cells, establishing a novel negative feedback loop that regulates RELA activation by TNF[alpha]. Additionally, we characterised the NF-[kappa]B pathway in PDAC cells, identifying how NF-[kappa]B/I[kappa]B proteins genetically and physically interact with RELA in the absence or presence of TNF[alpha]. Taken together, we provide computational and experimental support for interdependence between the F-actin network and the NF-[kappa]B pathway with RELA translocation dynamics in PDAC.
ISSN:2050-084X
2050-084X
DOI:10.7554/eLife.86042