Signal transduction through decay-accelerating factor. Interaction of glycosyl-phosphatidylinositol anchor and protein tyrosine kinases p56lck and p59fyn 1.
Decay-accelerating factor (DAF or CD55) is a 70-kDa glycosyl-phosphatidylinositol (GPI)-anchored protein that protects cells from complement-mediated lysis by either preventing the formation of or dissociating C3 convertases. Cross-linking of DAF on human peripheral T cells by polyclonal antibodies has previously been reported to lead to lymphocyte proliferation. Two mAb, both mapping to the third short consensus repeat region of DAF, were able to trigger proliferation of human peripheral T cells. To determine the role of the GPI anchor in cell activation, we transfected EL-4 murine thymoma cells with cDNA encoding either DAF or a transmembrane form of DAF (DAF-TM). The DAF-transfected cells were able to transduce late activation events as evidenced by IL-2 production, whereas DAF-TM transfected cells were unable to do so. The GPI-anchored DAF was able to transduce early activation events leading to the tyrosine phosphorylation of a 40-kDa protein and several proteins in the 85-95 kDa range--an event absent in DAF-TM-transfected cells. Furthermore, anti-DAF immunoprecipitates of DAF-transfected cells contain tyrosine kinase activity leading to the phosphorylation of 40-, 56-60-, and 85-kDa proteins, whereas anti-DAF immunoprecipitates of DAF-TM-transfected cells did not have an associated kinase activity. Both p56lck and p59fyn were associated with DAF in DAF-transfected EL-4 cells. In HeLa cells transfected with fyn, DAF associated with p59fyn. This complex of DAF with src family protein tyrosine kinases requires the GPI anchor and suggests a pathway for signaling through GPI-anchored membrane proteins.
Shenoy-Scaria
A M
AM
Department of Pathology, Washington University School of Medicine, St. Louis, MO 63110.
Kwong
J
J
Fujita
T
T
Olszowy
M W
MW
Shaw
A S
AS
Lublin
D M
DM
D015703:Antigens, CD / Q000502:physiology*; D018958:CD55 Antigens; D017261:Glycosylphosphatidylinositols / Q000502:physiology*; D006801:Humans; D066298:In Vitro Techniques; D008213:Lymphocyte Activation; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D008562:Membrane Glycoproteins / Q000502:physiology*; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051076:Proto-Oncogene Proteins c-fyn; D015398:Signal Transduction; D013601:T-Lymphocytes / Q000502:physiology*; D014162:Transfection
D015703:Antigens, CD; D018958:CD55 Antigens; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D011518:Proto-Oncogene Proteins; D011505:Protein-Tyrosine Kinases; C494957:FYN protein, human; D019860:Lymphocyte Specific Protein Tyrosine Kinase p56(lck); D051076:Proto-Oncogene Proteins c-fyn