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.
Cai
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Department of Medicine, Section of Hematology and Oncology, Boston Medical Center, MA 02118, USA.
Clayton
L K
LK
Smolyar
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A
Lerner
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D016475:3T3 Cells; D048868:Adaptor Proteins, Signal Transducing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002448:Cell Adhesion / Q000276:immunology; D018797:Cell Cycle Proteins / Q000737:chemistry; D050719:Crk-Associated Substrate Protein; D016207:Cytokines / Q000502:physiology*; D020662:Guanine Nucleotide Exchange Factors; D007375:Interleukin-1 / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D011485:Protein Binding / Q000276:immunology; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D050717:Retinoblastoma-Like Protein p130; D017386:Sequence Homology, Amino Acid; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism; D013950:Thymus Gland / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000502:physiology; D014443:Tyrosine / Q000378:metabolism; D020666:ras-GRF1
D048868:Adaptor Proteins, Signal Transducing; C493842:Bcar1 protein, mouse; C120867:Bcar3 protein, mouse; D018797:Cell Cycle Proteins; D050719:Crk-Associated Substrate Protein; D016207:Cytokines; D020662:Guanine Nucleotide Exchange Factors; D007375:Interleukin-1; D010446:Peptide Fragments; D010750:Phosphoproteins; D011506:Proteins; D050717:Retinoblastoma-Like Protein p130; D014409:Tumor Necrosis Factor-alpha; D020666:ras-GRF1; D014443:Tyrosine