AND-34, a novel p130Cas-binding thymic stromal cell protein regulated by adhesion and inflammatory cytokines.
We have characterized a novel cDNA whose steady state mRNA levels rise in the thymus 2 to 6 h following the induction of CD4+CD8+ thymocyte apoptosis by in vivo cross-linking of CD3 epsilon. This cDNA, AND-34-1, contains an open reading frame (ORF) encoding a protein with an amino-terminal Src homology ... 2 (SH2) domain and a carboxyl-terminal domain homologous to GDP-exchange factors (GEFs). Northern analysis demonstrates widespread expression of the AND-34 gene. Anti-CD3 epsilon treatment induces up-regulation of the AND-34 mRNA levels in total thymic RNA but not in RNA from purified thymocytes, suggesting that this transcript is derived from a thymic stromal cell population. IL-1 and TNF increase AND-34 transcript levels in thymic cortical reticular, thymic nurse, and fibroblast cell lines. In the thymic cortical reticular cell line, IL-1 and TNF induce a protein of the predicted 93-kDa size reactive with anti-AND-34 peptide antisera. Fifteen minutes of serum stimulation of vanadate-pretreated AND-34-1-transfected NIH3T3 fibroblasts induces tyrosine phosphorylation of AND-34 as well as coprecipitating 95-, 125-, and 130-kDa proteins. One of these tyrosine phosphorylated proteins is identified as p130Cas (Crk-associated substrate), a signaling molecule previously known to bind to a GDP-exchange factor (C3G) and inducibly associate with the focal adhesion complex. Consistent with such an association, AND-34 tyrosine phosphorylation is induced following adherence of trypsinized fibroblasts to fibronectin or poly-L -lysine-coated surfaces.
Mesh Terms:
3T3 Cells, Amino Acid Sequence, Animals, Base Sequence, Cell Adhesion, Cell Cycle Proteins, Crk-Associated Substrate Protein, Cytokines, Guanine Nucleotide Exchange Factors, Interleukin-1, Mice, Mice, Inbred C57BL, Mice, Transgenic, Molecular Sequence Data, Peptide Fragments, Phosphoproteins, Phosphorylation, Polymerase Chain Reaction, Precipitin Tests, Protein Binding, Proteins, Retinoblastoma-Like Protein p130, Sequence Homology, Amino Acid, Stromal Cells, Thymus Gland, Tumor Necrosis Factor-alpha, Tyrosine, ras-GRF1
3T3 Cells, Amino Acid Sequence, Animals, Base Sequence, Cell Adhesion, Cell Cycle Proteins, Crk-Associated Substrate Protein, Cytokines, Guanine Nucleotide Exchange Factors, Interleukin-1, Mice, Mice, Inbred C57BL, Mice, Transgenic, Molecular Sequence Data, Peptide Fragments, Phosphoproteins, Phosphorylation, Polymerase Chain Reaction, Precipitin Tests, Protein Binding, Proteins, Retinoblastoma-Like Protein p130, Sequence Homology, Amino Acid, Stromal Cells, Thymus Gland, Tumor Necrosis Factor-alpha, Tyrosine, ras-GRF1
J. Immunol.
Date: Aug. 15, 1999
PubMed ID: 10438950
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