Fc member of RI-Mediated Induction of TNF- alpha Gene Expression in the RBL-2H3 Mast Cell Line: Regulation by a Novel NF- Kappa B-Like Nuclear Binding Complex

Using rat basophilic leukemia (RBL-2H3) cells as a model, we investigated how aggregation of the high affinity receptor for IgE (Fc member of RI) regulates TNF- alpha gene expression. Antigenic stimulation of RBL-2H3 cells led to an increase in newly synthesized TNF- alpha mRNA that was dependent on...

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Veröffentlicht in:The Journal of immunology (1950) 1998-11, Vol.161 (9), p.4768-4776
Hauptverfasser: Pelletier, C, Varin-Blank, N, Rivera, J, Iannascoli, B, Marchand, F, David, B, Weyer, A, Blank, U
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container_end_page 4776
container_issue 9
container_start_page 4768
container_title The Journal of immunology (1950)
container_volume 161
creator Pelletier, C
Varin-Blank, N
Rivera, J
Iannascoli, B
Marchand, F
David, B
Weyer, A
Blank, U
description Using rat basophilic leukemia (RBL-2H3) cells as a model, we investigated how aggregation of the high affinity receptor for IgE (Fc member of RI) regulates TNF- alpha gene expression. Antigenic stimulation of RBL-2H3 cells led to an increase in newly synthesized TNF- alpha mRNA that was dependent on continuous receptor aggregation and did not require de novo protein synthesis. Kinetic analysis showed that maximal levels were achieved at 60 min and waned by 180 min of stimulation. Concomitant with the transcriptional activation of the TNF- alpha gene, the rapid appearance and disappearance of a previously uncharacterized nuclear NF- Kappa B DNA binding activity, comprised of two distinct protein complexes, were observed. These protein complexes bound to NF- Kappa B sites within the TNF- alpha gene and contained novel proteins (three species of M sub(r) between 90,000-110,000) distinct from the classical proteins in NF- Kappa B complexes. The induced NF- Kappa B binding activity required continuous receptor stimulation and induced NF- Kappa B-dependent reporter gene expression. Consistent with a role for the novel NF- Kappa B nuclear binding activity in TNF- alpha gene expression, deletion of several 5' Kappa B elements in the TNF- alpha promoter abolished all measurable Fc member of RI-dependent induction of a reporter construct. Pharmacologic agents that inhibited the NF- Kappa B binding activity also inhibited TNF- alpha mRNA expression. Our results demonstrate that a novel NF- Kappa B-like nuclear binding activity plays an important role in regulation of the rapid and transient transcriptional activation of the TNF- alpha gene via Fc member of RI.
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Consistent with a role for the novel NF- Kappa B nuclear binding activity in TNF- alpha gene expression, deletion of several 5' Kappa B elements in the TNF- alpha promoter abolished all measurable Fc member of RI-dependent induction of a reporter construct. Pharmacologic agents that inhibited the NF- Kappa B binding activity also inhibited TNF- alpha mRNA expression. 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title Fc member of RI-Mediated Induction of TNF- alpha Gene Expression in the RBL-2H3 Mast Cell Line: Regulation by a Novel NF- Kappa B-Like Nuclear Binding Complex
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