Identification and subcellular localization of proteins that are rapidly phosphorylated in tyrosine in response to colony-stimulating factor 1

A. Sengupta, W. K. Liu, Y. G. Yeung, D. C Y Yeung, A. R. Frackelton, E. R. Stanley

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Abstract

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.

Original languageEnglish (US)
Pages (from-to)8062-8066
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume85
Issue number21
StatePublished - 1988

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Macrophage Colony-Stimulating Factor
Tyrosine
Colony-Stimulating Factor Receptors
Proteins
Phosphorylation
Signal Transduction
Intercellular Signaling Peptides and Proteins
Phosphotyrosine
Growth Factor Receptors
Reducing Agents
Cell Surface Receptors
Serine
Macrophages
Temperature

ASJC Scopus subject areas

  • General
  • Genetics

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Identification and subcellular localization of proteins that are rapidly phosphorylated in tyrosine in response to colony-stimulating factor 1. / Sengupta, A.; Liu, W. K.; Yeung, Y. G.; Yeung, D. C Y; Frackelton, A. R.; Stanley, E. R.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 85, No. 21, 1988, p. 8062-8066.

Research output: Contribution to journalArticle

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AU - Sengupta, A.

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AU - Frackelton, A. R.

AU - Stanley, E. R.

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