TY - JOUR
T1 - Expression and function of TRAF-3 splice-variant isoforms in human lymphoma cell lines
AU - Gamper, Christopher
AU - Omene, Coral O.
AU - Van Eyndhoven, Winfried G.
AU - Glassman, Grace D.
AU - Lederman, Seth
N1 - Funding Information: This work was supported in part by NIH/NCI grant R0-1-CA55713 to Seth Lederman. Christopher Gamper is a M.D./Ph.D. student in the Integrated Program in Cellular, Molecular and Biophysical Studies supported by the Medical Scientist Training Program Grant No. 5-T32-GM07367. Dr. Winfried van Eyndhoven was a Ph.D. student in Biological Sciences supported by a training grant from Roche. The authors thank Dr. Rolf Freter and Dr. Padma Sridhar of Columbia University, New York for their helpful suggestions regarding RNAse protection assays and Dr. Ricardo Dalla Fevera of Columbia University for the kind gift of BJAB cells.
PY - 2001
Y1 - 2001
N2 - TRAF-3 gene products are signaling adaptor molecules required for lymphocytes to mediate T-dependent antibody responses in vivo. Previous work identified 8 splice-variant TRAF-3 mRNA species by RT-PCR that have the potential to encode novel isoforms, seven of which induce NF-κB activation when over-expressed in 293 cells. Here, their expression was characterized by RNAse protection assay, which showed the T cell line Jurkat D1.1 and the B cell lines BJAB, Daudi, and Raji each expressed mRNA encoding TRAF-3 splice-variants in approximately the same rank order (from highest to lowest); TRAF-3 Δ103aa, Δ83aa, full-length, Δ25aa, Δ52aa, Δ56aa, Δ27aa, and Δ221aa mRNA. The TRAF-3 Δ130aa mRNA was not detectable in any of the cell lines examined. The functional effect of over-expressing each TRAF-3 splice-variant on NF-κB activation was studied in the TRAF-5-responsive B cell line, BJAB. Of the seven TRAF-3 splice-variant isoforms that induce NF-κB activation in 293 cells, only TRAF-3 Δ27aa, Δ103aa, or Δ130aa induce NF-κB activation in BJAB cells. Together, these data indicate that a number of TRAF-3 splice-variant mRNAs are expressed and function in B and T lymphoma lines, which suggests that certain TRAF-3 splice-variant isoforms may participate in mediating the known functions of the TRAF-3 gene in lymphocytes.
AB - TRAF-3 gene products are signaling adaptor molecules required for lymphocytes to mediate T-dependent antibody responses in vivo. Previous work identified 8 splice-variant TRAF-3 mRNA species by RT-PCR that have the potential to encode novel isoforms, seven of which induce NF-κB activation when over-expressed in 293 cells. Here, their expression was characterized by RNAse protection assay, which showed the T cell line Jurkat D1.1 and the B cell lines BJAB, Daudi, and Raji each expressed mRNA encoding TRAF-3 splice-variants in approximately the same rank order (from highest to lowest); TRAF-3 Δ103aa, Δ83aa, full-length, Δ25aa, Δ52aa, Δ56aa, Δ27aa, and Δ221aa mRNA. The TRAF-3 Δ130aa mRNA was not detectable in any of the cell lines examined. The functional effect of over-expressing each TRAF-3 splice-variant on NF-κB activation was studied in the TRAF-5-responsive B cell line, BJAB. Of the seven TRAF-3 splice-variant isoforms that induce NF-κB activation in 293 cells, only TRAF-3 Δ27aa, Δ103aa, or Δ130aa induce NF-κB activation in BJAB cells. Together, these data indicate that a number of TRAF-3 splice-variant mRNAs are expressed and function in B and T lymphoma lines, which suggests that certain TRAF-3 splice-variant isoforms may participate in mediating the known functions of the TRAF-3 gene in lymphocytes.
KW - Cell surface molecules
KW - Human
KW - Molecular biology
KW - Signal transduction
KW - T cell-B cell collaboration
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U2 - https://doi.org/10.1016/S0198-8859(01)00284-1
DO - https://doi.org/10.1016/S0198-8859(01)00284-1
M3 - Article
C2 - 11600226
VL - 62
SP - 1167
EP - 1177
JO - Human Immunology
JF - Human Immunology
SN - 0198-8859
IS - 10
ER -