Fold-Change Detection of NF-?B at Target Genes with Different Transcript Outputs.

ABSTRACT: The transcription factor nuclear factor (NF)-?B promotes inflammatory and stress-responsive gene transcription across a range of cell types in response to the cytokine tumor necrosis factor (TNF). Although NF-?B signaling exhibits significant variability across single cells, some target genes supporting high levels of TNF-inducible transcription exhibit fold-change detection of NF-?B, which may buffer against stochastic variation in signaling molecules. It is unknown whether fold-change detection is maintained at NF-?B target genes with low levels of TNF-inducible transcription, for which stochastic promoter events may be more pronounced. Here, we used a microfluidic cell-trapping device to measure how TNF-induced activation of NF-?B controls transcription in single Jurkat T cells at the promoters of integrated HIV and the endogenous cytokine gene IL6, which produce only a few transcripts per cell. We tracked TNF-stimulated NF-?B RelA nuclear translocation by live-cell imaging and then quantified transcript number by RNA FISH in the same cell. We found that TNF-induced transcript abundance at 2 h for low- and high-abundance target genes correlates with similar strength with the fold change in nuclear NF-?B. A computational model of TNF-NF-?B signaling, which implements fold-change detection from competition for binding to ?B motifs, could reproduce fold-change detection across the experimentally measured range of transcript outputs. However, multiple model parameters affecting transcription had to be simultaneously varied across promoters to maintain fold-change detection while also matching other trends in the single-cell data for low-abundance transcripts. Our results suggest that cells use multiple biological mechanisms to tune transcriptional output while maintaining robustness of NF-?B fold-change detection.

SUBMITTER: Wong VC 

PROVIDER: S-EPMC6382958 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature