J Biol Chem. 1994 Sep 9;269(36):22492-5.

Aggregation of the intracellular domain of the type 1 tumor necrosis factor receptor defined by the two-hybrid system.External

Song, H. Y., Dunbar, J. D., Donner, D. B.,
--- - Department of Physiology and Biophysics, Indiana University School of Medicine, Indianapolis 46202.
The yeast-based two hybrid system has been used to determine whether oligomerization of the intracellular domain of the 55-kDa type 1 tumor necrosis factor (TNF) receptor may occur during TNF action. This assay depends upon reconstitution of the function of the GAL4 transcriptional activator through interaction of a protein fused to the GAL4 DNA binding domain with a protein fused to the transcriptional activation domain of GAL4. Fusion of the type 1 TNF receptor intracellular domain with the DNA binding domain and the transactivation domain of GAL4 led to activation of the lacZ indicator gene, demonstrating interaction of the receptor intracellular domain with itself. A HeLa cell cDNA library was searched for proteins that interact with the intracellular domain of the type 1 TNF receptor. A protein corresponding to amino acids 329-426 in the type 1 TNF receptor intracellular domain was identified by this screen. The aggregation domain was further defined by testing the ability of deletion mutants of the type 1 TNF receptor intracellular region to interact with the complete intracellular domain. These experiments map the aggregation domain to a sequence of amino acids previously shown to be responsible for mediating TNF-induced cytotoxicity. These results suggest that aggregation of type 1 TNF receptor intracellular domains may be important in TNF signal transduction.
PMID: 8077196External
Arrow2 In vitro interaction Arrow2 In vivo interaction Arrow2 Characterization Arrow2 Functional role Arrow2 top
In vitro interaction
  DD1 DD2 Reference
Family DD1 DD2 Method Species Region Expression Species Region Expression
DD TNFR1 Link TNFR1 Yeast two-hybrid Not specified 204-426 Yeast Not specified 204-426 Yeast 8077196
DD TNFR1 Link TNFR1 Yeast two-hybrid Not specified 204-426 Yeast Not specified 329-426 Yeast 8077196
(Link: click this icon to show interactions only between the two corresponding DDs)