CD30 Contains Two Binding Sites with Different Specificities for Members of the Tumor Necrosis Factor Receptor-associated Factor Family of Signal Transducing Proteins

Abstract

CD30 is a member of the tumor necrosis factor (TNF) receptor family of proteins. CD30 can regulate proliferation of lymphocytes and may also play an important role in human immunodeficiency virus replication. However, little is known about CD30 signal transduction. We performed a yeast two-hybrid library screen with the cytoplasmic domain of CD30 and isolated multiple independent cDNAs encoding human tumor necrosis factor receptor-associated factor (TRAF) 1, TRAF2, and CRAF1 (TRAF3). The ability of TRAF1, TRAF2, and CRAF1 to associate with CD30 was confirmed using an in vitro coprecipitation assay, further demonstrating that the interaction was specific and direct. The TRAF-binding domain of CD30 was mapped to the COOH-terminal 36 amino acid residues, which contained two independent binding sites. CRAF1 bound only a single site, which contained the sequence PEQET, whereas TRAF1 and TRAF2 were capable of binding to either the PEQET site or an additional downstream domain. These data indicate that the TRAF protein binding pattern of CD30 differs from other TNF receptor family members and suggest that signaling specificity through TNF receptor family proteins may be achieved through differences in their abilities to bind TRAF proteins.

Glucocorticoid-Induced TNF Receptor, a Costimulatory Receptor on Naive and Activated T Cells, Uses TNF Receptor-Associated Factor 2 in a Novel Fashion as an Inhibitor of NF-κB Activation

“…Peritoneal macrophages from 129/SvJ mice (a kind gift of Dr. R. Schreiber, Washington University, School of Medicine) and activated DO11.10 splenocytes were used to generate cDNA for PCR amplification of full-length GITR-L and GITR, respectively, which were cloned into pcDNA3.1 (Invitrogen Life Technologies). TRAF2 was subcloned into pcDNA3 (33). Corresponding alanine substitution and deletion mutations of pAS1 [GITRcp] and pcDNA3.1 [GITR] were generated using the QuikChange mutagenesis system (Stratagene) and the following sense primers and complementary oligonucleotides and verified by sequencing: 202EDϾ2A, 5Ј-GGAAGCTG CAAGCCGCGGCAGCTGACAACTGC-3Ј; 211STOP, 5Ј-CTTCTGTCT GCTCCCGCGGTTACTACTAAGGGAACTGGAAGC-3Ј; 211EEEϾ3A 5Ј-GCTTCCAGTTCCCTGCCGCGGCCCGCGGGGAGCAGACAGAAG-3Ј; 219EEϾ2A 5Ј-GGGGAGCAGACCGCGGCAAAGTGTCATCT GGGGGG-3Ј.…”

TCR-Independent CD30 Signaling Selectively Induces IL-13 Production Via a TNF Receptor-Associated Factor/p38 Mitogen-Activated Protein Kinase-Dependent Mechanism

“…In transfected COS or 293 cells, CD30 associates with TRAF1, TRAF2, and TRAF3 (15,16). There are two conserved C-terminal TRAF binding sites in the CD30 cytoplasmic tail, which both can bind TRAF1 and TRAF2, whereas TRAF3 can only bind to the upstream site (17). Both conserved TRAF binding sites in CD30 are able to mediate NF-B activation (18).…”

TNF Receptor-Associated Factor (TRAF) Signaling Network in CD4⁺ T-Lymphocytes

“…When bound by specific ligands, these costimulatory TNFRs recruit TRAF1, TRAF2, TRAF3, and TRAF5 (Figs. 3 and 4) (Gedrich et al 1996;Lee et al 1996;Aizawa et al 1997;Boucher et al 1997;Duckett et al 1997 (Rothe et al 1994;Boucher et al 1997). DR3 recruits TRAF2 via the TNFR-associated death domain protein (TRADD) (Park et al 2000).…”

“…TRAF2 is recruited to 4-1BB (Arch and Thompson 1998;Jang et al 1998;Saoulli et al 1998), CD27 (Akiba et al 1998;Gravestein et al 1998;Yamamoto et al 1998), CD30 (Gedrich et al 1996;Lee et al 1996;Harlin et al 2002), DR3 (Chinnaiyan et al 1996;Pobezinskaya et al 2011), GITR (Kwon et al 1999;Esparza and Arch 2005), HVEM (Hsu et al 1997;Marsters et al 1997;Cheung et al 2009), OX40 (Arch and Thompson 1998;Kawamata et al 1998), and TNFR2 (Rothe et al 1994).…”

“…TRAF1 is recruited to 4-1BB (Arch and Thompson 1998;Jang et al 1998;Saoulli et al 1998), CD30 (Gedrich et al 1996;Lee et al 1996), GITR (Kwon et al 1999), HVEM (Marsters et al 1997), OX40 (Kawamata et al 1998), and TNFR2 (Rothe et al 1994). …”