caa a22 4500 606175512 CHVBK 20210128100743.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s00018-015-1850-1 doi (NATIONALLICENCE)springer-10.1007/s00018-015-1850-1 Species differential regulation of COX2 can be described by an NFκB-dependent logic AND gate [Elektronische Daten] [Lan Nguyen, Miguel Cavadas, Boris Kholodenko, Till Frank, Alex Cheong] Cyclooxygenase 2 (COX2), a key regulatory enzyme of the prostaglandin/eicosanoid pathway, is an important target for anti-inflammatory therapy. It is highly induced by pro-inflammatory cytokines in a Nuclear factor kappa B (NFκB)-dependent manner. However, the mechanisms determining the amplitude and dynamics of this important pro-inflammatory event are poorly understood. Furthermore, there is significant difference between human and mouse COX2 expression in response to the inflammatory stimulus tumor necrosis factor alpha (TNFα). Here, we report the presence of a molecular logic AND gate composed of two NFκB response elements (NREs) which controls the expression of human COX2 in a switch-like manner. Combining quantitative kinetic modeling and thermostatistical analysis followed by experimental validation in iterative cycles, we show that the human COX2 expression machinery regulated by NFκB displays features of a logic AND gate. We propose that this provides a digital, noise-filtering mechanism for a tighter control of expression in response to TNFα, such that a threshold level of NFκB activation is required before the promoter becomes active and initiates transcription. This NFκB-regulated AND gate is absent in the mouse COX2 promoter, most likely contributing to its differential graded response in promoter activity and protein expression to TNFα. Our data suggest that the NFκB-regulated AND gate acts as a novel mechanism for controlling the expression of human COX2 to TNFα, and its absence in the mouse COX2 provides the foundation for further studies on understanding species-specific differential gene regulation. The Author(s), 2015 NFκB signaling nationallicence COX2 nationallicence Switch-like gene regulation nationallicence Logic AND gate nationallicence Kinetic modeling nationallicence COX1 : Cyclooxygenase 1 nationallicence COX2 : Cyclooxygenase 2 nationallicence hCOX2 : Human cyclooxygenase 2 nationallicence mCOX2 : Mouse cyclooxygenase 2 nationallicence TNFα : Tumor necrosis factor alpha nationallicence NFκB : Nuclear factor kappa B nationallicence NRE : NFκB response element nationallicence Gluc : Gaussia luciferase nationallicence pGluc : Plasmid encoding Gluc nationallicence CRE : Cis-regulatory elements nationallicence Nguyen Lan Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut Cavadas Miguel Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut Kholodenko Boris Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut Frank Till Center for the Ecological Study of Perception and Action, University of Connecticut, 06269, Storrs, CT, USA aut Cheong Alex Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut Cellular and Molecular Life Sciences Springer Basel 72/12(2015-06-01), 2431-2443 1420-682X 72:12<2431 2015 72 18 https://doi.org/10.1007/s00018-015-1850-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00018-015-1850-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nguyen Lan Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut NATIONALLICENCE 700 1- Cavadas Miguel Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut NATIONALLICENCE 700 1- Kholodenko Boris Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut NATIONALLICENCE 700 1- Frank Till Center for the Ecological Study of Perception and Action, University of Connecticut, 06269, Storrs, CT, USA aut NATIONALLICENCE 700 1- Cheong Alex Systems Biology Ireland, University College Dublin, Dublin 4, Ireland aut NATIONALLICENCE 773 0- Cellular and Molecular Life Sciences Springer Basel 72/12(2015-06-01), 2431-2443 1420-682X 72:12<2431 2015 72 18