title: Nuclear translocation of STAT3 and NF-κB are independent of each other but NF-κB supports expression and activation of STAT3.
abstract: NF-κB and STAT3 are essential transcription factors in immunity and act at the interface of the transition from chronic inflammation to cancer. Different functional crosstalks between NF-κB and STAT3 have been recently described arguing for a direct interaction of both proteins. During a systematic analysis of NF-κB/STAT3 crosstalk we observed that appearance of the subcellular distribution of NF-κB and STAT3 in immunofluorescence heavily depends on the fixation procedure. Therefore, we established an optimized fixation protocol for the reliable simultaneous analysis of the subcellular distributions of both transcription factors. Using this protocol we found that cytokine-induced nuclear accumulation of NF-κB or STAT3 did not alter the subcellular distribution of the other transcription factor. Both knockout and overexpression of STAT3 does not have any major effect on canonical TNFα-NF-κB signalling in MEF or HeLa cells. Similarly, knockout of p65 did not alter nuclear accumulation of STAT3 in response to IL-6. However, p65 expression correlates with elevated total cellular levels of STAT3 and STAT1 and supports activation of these transcription factors. Our findings in MEF cells argue against a direct physical interaction of free cellular NF-κB and STAT3 but point to more intricate functional interactions.
authors.affiliation: Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Antons
authors.initials: A
authors.lastname: Martincuks
authors.affiliation: Systems Biology of Biochemical Signalling, Laboratory of Modelling in Biology and Medicine, Institute of Fundamental Technological Research, Polish Academy of Sciences, Pawinskiego 5b, 02-106 Warszawa, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Andryka
authors.affiliation: Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Andrea
authors.initials: A
authors.lastname: Küster
authors.affiliation: Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany.
authors.forename: Hildegard
authors.initials: H
authors.lastname: Schmitz-Van de Leur
authors.affiliation: Systems Biology of Biochemical Signalling, Laboratory of Modelling in Biology and Medicine, Institute of Fundamental Technological Research, Polish Academy of Sciences, Pawinskiego 5b, 02-106 Warszawa, Poland.
authors.forename: Michal
authors.initials: M
authors.lastname: Komorowski
authors.affiliation: Institute of Biochemistry and Molecular Biology, RWTH Aachen University, Pauwelsstraße 30, 52074 Aachen, Germany. Electronic address: mueller-newen@rwth-aachen.de.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Müller-Newen
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006367:HeLa Cells; D006801:Humans; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D050794:STAT1 Transcription Factor / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D050799:STAT5 Transcription Factor / Q000378:metabolism; D051996:Transcription Factor RelA / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D016328:NF-kappa B; D050794:STAT1 Transcription Factor; D050796:STAT3 Transcription Factor; D050799:STAT5 Transcription Factor; D051996:Transcription Factor RelA; D014409:Tumor Necrosis Factor-alpha
keywords: Crosstalk; NF-κB; Nuclear translocation; STAT3; Signal transduction