02597nas a2200265   4500008004100000245011200041210007000153260000900223300000800232490000600240520167600246100001901922700002501941700001801966700002301984700003002007700002502037700001902062700001902081700001702100700001802117700002402135700002002159856015202179       2014                        eng d00aNOTCH1 Can Initiate NF-κB Activation via Cytosolic Interactions with Components of the T Cell Signalosome.0 aNOTCH1 Can Initiate NFκB Activation via Cytosolic Interactions w  c2014  a2490 v53 a<p>T cell stimulation requires the input and integration of external signals. Signaling through the T cell receptor (TCR) is known to induce formation of the membrane-tethered CBM complex, comprising CARMA1, BCL10, and MALT1, which is required for TCR-mediated NF-κB activation. TCR signaling has been shown to activate NOTCH proteins, transmembrane receptors also implicated in NF-κB activation. However, the link between TCR-mediated NOTCH signaling and early events leading to induction of NF-κB activity remains unclear. In this report, we demonstrate a novel cytosolic function for NOTCH1 and show that it is essential to CBM complex formation. Using a model of skin allograft rejection, we show in vivo that NOTCH1 acts in the same functional pathway as PKCθ, a T cell-specific kinase important for CBM assembly and classical NF-κB activation. We further demonstrate in vitro NOTCH1 associates physically with PKCθ and CARMA1 in the cytosol. Unexpectedly, when NOTCH1 expression was abrogated using RNAi approaches, interactions between CARMA1, BCL10, and MALT1 were lost. This failure in CBM assembly reduced inhibitor of kappa B alpha phosphorylation and diminished NF-κB-DNA binding. Finally, using a luciferase gene reporter assay, we show the intracellular domain of NOTCH1 can initiate robust NF-κB activity in stimulated T cells, even when NOTCH1 is excluded from the nucleus through modifications that restrict it to the cytoplasm or hold it tethered to the membrane. Collectively, these observations provide evidence that NOTCH1 may facilitate early events during T cell activation by nucleating the CBM complex and initiating NF-κB signaling.</p>1 aShin, Hyun, Mu1 aTilahun, Mulualem, E1 aCho, Ok, Hyun1 aChandiran, Karthik1 aKuksin, Christina, Arieta1 aKeerthivasan, Shilpa1 aFauq, Abdul, H1 aGolde, Todd, E1 aMiele, Lucio1 aThome, Margot1 aOsborne, Barbara, A1 aMinter, Lisa, M  uhttps://www.umass.edu/veterinary-animal-sciences/research-faculty/publications/notch1-can-initiate-nf-kb-activation-via-cytosolic-interactions-with