TY - JOUR
T1 - Identification and subcellular localization of proteins that are rapidly phosphorylated in tyrosine in response to colony-stimulating factor 1
AU - Sengupta, A.
AU - Liu, W. K.
AU - Yeung, Y. G.
AU - Yeung, D. C.Y.
AU - Frackelton, A. R.
AU - Stanley, E. R.
PY - 1988
Y1 - 1988
N2 - To investigate growth factor-mediated signal transduction, we have studied phosphorylation events that take place within seconds of the binding of colony-stimulating factor 1 (CSF-1) to its cell-surface receptor. CSF-1 stimulated rapid tyrosine phosphorylation of cellular proteins in murine BAC1.2F5 macrophages at 37°C and 4°C. The pattern of CSF-1-stimulated tyrosine phosphorylation of at least 15 different proteins at both temperatures was similar and unchanged by treatment of the lysate with reducing agent. With the exception of the 185-kDa CSF-1 receptor, a 260-kDa protein and a 133-kDa protein, the proteins were predominantly cytoplasmic. At 37°C, all the proteins were phosphorylated within 30 sec of addition of growth factor. At 4°C, CSF-1 receptor sites were saturated after 2 min of incubation in the presence of high concentrations of CSF-1 and differences in the order of appearance of phosphorylated proteins were observed: 185 kDa (CSF-1 receptor) (by 2 min); 99 kDa (by 4 min); 125 kDa (by 10 min); 61 kDa (by 30 min); and 260 kDa, 84 kDa, and 41 kDa (by 180 min). In addition to stimulating the phosphorylation of these proteins in tyrosine, CSF-1 caused dephosphorylation of phosphorylated serine residues on the receptor. As neither CSF-1 nor its receptor is internalized at 4°C, analysis of these early reactions and the phosphotyrosine-containing proteins in intact cells under these conditions should lead to an understanding of the early events in growth factor receptor-mediated signal transduction.
AB - To investigate growth factor-mediated signal transduction, we have studied phosphorylation events that take place within seconds of the binding of colony-stimulating factor 1 (CSF-1) to its cell-surface receptor. CSF-1 stimulated rapid tyrosine phosphorylation of cellular proteins in murine BAC1.2F5 macrophages at 37°C and 4°C. The pattern of CSF-1-stimulated tyrosine phosphorylation of at least 15 different proteins at both temperatures was similar and unchanged by treatment of the lysate with reducing agent. With the exception of the 185-kDa CSF-1 receptor, a 260-kDa protein and a 133-kDa protein, the proteins were predominantly cytoplasmic. At 37°C, all the proteins were phosphorylated within 30 sec of addition of growth factor. At 4°C, CSF-1 receptor sites were saturated after 2 min of incubation in the presence of high concentrations of CSF-1 and differences in the order of appearance of phosphorylated proteins were observed: 185 kDa (CSF-1 receptor) (by 2 min); 99 kDa (by 4 min); 125 kDa (by 10 min); 61 kDa (by 30 min); and 260 kDa, 84 kDa, and 41 kDa (by 180 min). In addition to stimulating the phosphorylation of these proteins in tyrosine, CSF-1 caused dephosphorylation of phosphorylated serine residues on the receptor. As neither CSF-1 nor its receptor is internalized at 4°C, analysis of these early reactions and the phosphotyrosine-containing proteins in intact cells under these conditions should lead to an understanding of the early events in growth factor receptor-mediated signal transduction.
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U2 - 10.1073/pnas.85.21.8062
DO - 10.1073/pnas.85.21.8062
M3 - Article
C2 - 2460861
AN - SCOPUS:0000172885
SN - 0027-8424
VL - 85
SP - 8062
EP - 8066
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 21
ER -