NF-κB modifies the mammalian circadian clock through interaction with the core clock protein BMAL1

• Published in: PLoS genetics 2021-11, Vol.17 (11), p.e1009933-e1009933
• Main Authors: Shen, Yang, Endale, Mehari, Wang, Wei, Morris, Andrew R, Francey, Lauren J, Harold, Rachel L, Hammers, David W, Huo, Zhiguang, Partch, Carrie L, Hogenesch, John B, Wu, Zhao-Hui, Liu, Andrew C
• Format: Article
• Language: English
• Subjects: Animals; ARNTL Transcription Factors - genetics; Biology and Life Sciences; Bioluminescence; BMAL1 protein; Calcium-Binding Proteins - genetics; Cell Line, Tumor; Chromatin; Circadian Clocks - genetics; Circadian rhythm; Circadian Rhythm - genetics; Circadian rhythms; Clock gene; CLOCK Proteins - genetics; Cryptochromes; Cryptochromes - genetics; Gene expression; Gene Expression Regulation, Developmental - genetics; Humans; Immunoprecipitation; Independent sample; Inflammation; Inflammation - genetics; Inflammation - pathology; Kinases; Locomotor activity; Mammals; NF-kappa B - genetics; NF-κB protein; Phenotypes; Physiology; RelA protein; Research and analysis methods; Suprachiasmatic nucleus; Suprachiasmatic Nucleus - metabolism; Transcription Factor RelA - genetics; Transcription factors
• ISSN: 1553-7404; 1553-7390; 1553-7404
• DOI: 10.1371/journal.pgen.1009933

In mammals, the circadian clock coordinates cell physiological processes including inflammation. Recent studies suggested a crosstalk between these two pathways. However, the mechanism of how inflammation affects the clock is not well understood. Here, we investigated the role of the proinflammatory transcription factor NF-κB in regulating clock function. Using a combination of genetic and pharmacological approaches, we show that perturbation of the canonical NF-κB subunit RELA in the human U2OS cellular model altered core clock gene expression. While RELA activation shortened period length and dampened amplitude, its inhibition lengthened period length and caused amplitude phenotypes. NF-κB perturbation also altered circadian rhythms in the master suprachiasmatic nucleus (SCN) clock and locomotor activity behavior under different light/dark conditions. We show that RELA, like the clock repressor CRY1, repressed the transcriptional activity of BMAL1/CLOCK at the circadian E-box cis-element. Biochemical and biophysical analysis showed that RELA binds to the transactivation domain of BMAL1. These data support a model in which NF-kB competes with CRY1 and coactivator CBP/p300 for BMAL1 binding to affect circadian transcription. This is further supported by chromatin immunoprecipitation analysis showing that binding of RELA, BMAL1 and CLOCK converges on the E-boxes of clock genes. Taken together, these data support a significant role for NF-κB in directly regulating the circadian clock and highlight mutual regulation between the circadian and inflammatory pathways.