TY - JOUR
T1 - TRAF-3 interacts with p62 nucleoporin, a component of the nuclear pore central plug that binds classical NLS-containing import complexes
AU - Gamper, C.
AU - Van Eyndhoven, W. G.
AU - Schweiger, E.
AU - Mossbacher, M.
AU - Koo, B.
AU - Lederman, S.
N1 - Funding Information:
This work was supported in part by NIH/NCI grant R0-1-CA55713 to Seth Lederman. Christopher Gamper is a MD/PhD student in the Integrated Program in Cellular, Molecular and Biophysical Studies supported by the Medical Scientist Training Program Grant ♯ 5-T32-GM07367. Dr Winfried van Eyndhoven was a PhD student in Biological Sciences supported by a training grant from Roche. The authors thank Dr Dimitris Thanos of Columbia University, New York for pCNDA3/p65(RelA), Dr Stephen Goff of Columbia University for pCGN, and Dr Paul B. Rothman of Columbia University for B2WTx3Luc.
PY - 2000/1/1
Y1 - 2000/1/1
N2 - The TRAF-3 gene encodes a number of splice-variant isoforms that function as adapter molecules in NF-κB signaling, in part by associating with the cytoplasmic tails of CD40 or other TNF-receptor (TNF-R) family members. To identify downstream molecules in TRAF-3 signaling, a yeast two-hybrid library was screened with a full-length TRAF-3 construct. Nine independent TRAF-3 interacting clones encoded fragments of p62 Nucleoporin (p62), a 522 amino acid (aa) component of the nuclear pore central plug, that is known to bind karyopherin-β/classical-NLS import factor complexes. The interaction of p62 with TRAF-3 was specific, since p62 failed to interact with TRAF-2, -4, -5, or -6. Deletional analysis in yeast revealed that the p62:TRAF-3 interaction is mediated by a p62 carboxy (C)-terminal coiled-coil domain. In human 293 T cells, recombinant TRAF-3 or p62 specifically co-immunoprecipitates the other species. In addition, endogenous p62 co-precipitates over-expressed TRAF-3. The functional effects of over-expressing a TRAF-3 binding fragment, p62(aa 336-522) were studied on NF-κB-dependent, or control STAT1-dependent reporter activity in 293 T cells, either resting or after stimulation by CD40 or IFN-γ, respectively. Over-expression of p62(aa 336-522) induces NF-κB activation in resting cells and augments CD40-induced NF-κB activation, but has no effect on control STAT1 reporter activity, either at baseline or after IFN-γ induction. The finding that TRAF-3 binds p62, suggests that TRAF-3 may serve as an adapter molecule at the nuclear membrane, in addition to its known adapter function at the plasma membrane. (C) 2000 Elsevier Science Ltd. All rights reserved.
AB - The TRAF-3 gene encodes a number of splice-variant isoforms that function as adapter molecules in NF-κB signaling, in part by associating with the cytoplasmic tails of CD40 or other TNF-receptor (TNF-R) family members. To identify downstream molecules in TRAF-3 signaling, a yeast two-hybrid library was screened with a full-length TRAF-3 construct. Nine independent TRAF-3 interacting clones encoded fragments of p62 Nucleoporin (p62), a 522 amino acid (aa) component of the nuclear pore central plug, that is known to bind karyopherin-β/classical-NLS import factor complexes. The interaction of p62 with TRAF-3 was specific, since p62 failed to interact with TRAF-2, -4, -5, or -6. Deletional analysis in yeast revealed that the p62:TRAF-3 interaction is mediated by a p62 carboxy (C)-terminal coiled-coil domain. In human 293 T cells, recombinant TRAF-3 or p62 specifically co-immunoprecipitates the other species. In addition, endogenous p62 co-precipitates over-expressed TRAF-3. The functional effects of over-expressing a TRAF-3 binding fragment, p62(aa 336-522) were studied on NF-κB-dependent, or control STAT1-dependent reporter activity in 293 T cells, either resting or after stimulation by CD40 or IFN-γ, respectively. Over-expression of p62(aa 336-522) induces NF-κB activation in resting cells and augments CD40-induced NF-κB activation, but has no effect on control STAT1 reporter activity, either at baseline or after IFN-γ induction. The finding that TRAF-3 binds p62, suggests that TRAF-3 may serve as an adapter molecule at the nuclear membrane, in addition to its known adapter function at the plasma membrane. (C) 2000 Elsevier Science Ltd. All rights reserved.
KW - Cell surface molecules
KW - Human
KW - Molecular biology
KW - Signal transduction
KW - T cell-B cell collaboration
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U2 - 10.1016/S0161-5890(00)00015-8
DO - 10.1016/S0161-5890(00)00015-8
M3 - Article
C2 - 10781837
AN - SCOPUS:0034066940
SN - 0161-5890
VL - 37
SP - 73
EP - 84
JO - Molecular Immunology
JF - Molecular Immunology
IS - 1-2
ER -