scholarly journals Antipeptide antiserum identifies a widely distributed cellular tyrosine kinase related to but distinct from the c-fps/fes-encoded protein.

1986 ◽  
Vol 6 (4) ◽  
pp. 1065-1073 ◽  
Author(s):  
R A Feldman ◽  
J P Tam ◽  
H Hanafusa
Keyword(s):  
Tyrosine Kinase ◽  
Mammalian Cells ◽  
Adult Life ◽  
Kinase Domain ◽  
Cellular Protein ◽  
Antigenic Determinants ◽  
Protein Kinase Activity ◽  
Normal Cells ◽  
Cellular Homolog ◽  
Sarcoma Virus

We raised antibodies directed against a synthetic peptide representing an amino acid sequence of the conserved kinase domain of the transforming protein of Fujinami sarcoma virus (FSV) (P140). The antiserum obtained specifically recognized FSV-P140 and its cellular homolog and in addition, it recognized a new cellular protein of 94,000 daltons (NCP94) in avian and mammalian cells. NCP94 was found to be associated with a cyclic nucleotide-independent protein kinase activity that was specific for tyrosine residues. Although NCP94 and FSV-P140 share antigenic determinants, NCP94 is not a cellular homolog of FSV-P140: NCP94 and the previously identified c-fps/fes product were different in their tryptic fingerprints and in their tissue specificities. Thus, the function of NCP94 in normal cells is probably different than that of the c-fps/fes product. NCP94 was expressed in every tissue and cell line that was examined. In chickens, NCP94 levels were highest during embryonic development and NCP94 expression was high in gizzard, brain, and spleen throughout embryonic and adult life. The universal expression of NCP94 suggests that this protein may be involved in an essential function of normal cells. NCP94 may be a new cellular tyrosine kinase of the src gene family.

Antipeptide antiserum identifies a widely distributed cellular tyrosine kinase related to but distinct from the c-fps/fes-encoded protein

1986 ◽  
Vol 6 (4) ◽  
pp. 1065-1073
Author(s):  
R A Feldman ◽  
J P Tam ◽  
H Hanafusa
Keyword(s):  
Tyrosine Kinase ◽  
Mammalian Cells ◽  
Adult Life ◽  
Kinase Domain ◽  
Cellular Protein ◽  
Antigenic Determinants ◽  
Protein Kinase Activity ◽  
Normal Cells ◽  
Cellular Homolog ◽  
Sarcoma Virus

We raised antibodies directed against a synthetic peptide representing an amino acid sequence of the conserved kinase domain of the transforming protein of Fujinami sarcoma virus (FSV) (P140). The antiserum obtained specifically recognized FSV-P140 and its cellular homolog and in addition, it recognized a new cellular protein of 94,000 daltons (NCP94) in avian and mammalian cells. NCP94 was found to be associated with a cyclic nucleotide-independent protein kinase activity that was specific for tyrosine residues. Although NCP94 and FSV-P140 share antigenic determinants, NCP94 is not a cellular homolog of FSV-P140: NCP94 and the previously identified c-fps/fes product were different in their tryptic fingerprints and in their tissue specificities. Thus, the function of NCP94 in normal cells is probably different than that of the c-fps/fes product. NCP94 was expressed in every tissue and cell line that was examined. In chickens, NCP94 levels were highest during embryonic development and NCP94 expression was high in gizzard, brain, and spleen throughout embryonic and adult life. The universal expression of NCP94 suggests that this protein may be involved in an essential function of normal cells. NCP94 may be a new cellular tyrosine kinase of the src gene family.

Towards a molecular description of cell transformation by avian sarcoma virus

1980 ◽  
Vol 210 (1180) ◽  
pp. 387-396 ◽  
Keyword(s):  
Protein Kinase ◽  
Protein Phosphorylation ◽  
Molecular Mass ◽  
Cellular Protein ◽  
Transformed Cells ◽  
Relative Molecular Mass ◽  
Protein Kinase Activity ◽  
Avian Sarcoma Virus ◽  
Sarcoma Virus ◽  
Avian Sarcoma

The avian sarcoma virus transforming gene product has been identified and partially purified from extracts of transformed cells. It is a phosphoprotein with a relative molecular mass of 60 000 (pp60 src ) with two major sites of phosphorylation. pp60 src appears to be a cyclic-AMP-independent protein kinase as judged by protein phosphorylation with partly purified fractions. The specificity of the phosphorylation observed was judged by inhibition with anti-pp60 src IgG but not by normal IgG and by the fact that the protein kinase activity isolated from ts transformation-mutant infected cells was more thermolabile than that from wild-type transformed cells, thus showing more directly the origin of the enzymic activity. A cellular protein substrate of pp60 src has been identified as a 34000 molecular mass protein. These data together suggest that protein phosphorylation by pp60 src may be a function of the molecule that plays a major role in transformation.

Amino acid substitutions sufficient to convert the nontransforming p60c-src protein to a transforming protein

1986 ◽  
Vol 6 (12) ◽  
pp. 4155-4160
Author(s):  
J Y Kato ◽  
T Takeya ◽  
C Grandori ◽  
H Iba ◽  
J B Levy ◽  
...  
Keyword(s):  
Amino Acid ◽  
Biological Activities ◽  
Tumor Formation ◽  
Protein Kinase Activity ◽  
Focus Formation ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Cellular Homolog ◽  
Sarcoma Virus ◽  
Hybrid Genes

We have previously shown that Rous sarcoma virus variants that carry the cellular homolog (c-src) of the viral src gene (v-src) do not transform chicken embryo fibroblasts. We also have shown that replacement of sequences upstream or downstream from the BglI site of the cellular src gene with the corresponding regions of v-src restored transforming activity to the hybrid genes. Since there are only six amino acid changes between p60c-src and p60v-src within the sequences upstream from BglI, we constructed chimeric molecules involving v-src and c-src to determine the effect of each amino acid substitution on the biological activities of the gene product. We found that the change from Thr to Ile at position 338 or the replacement of a fragment of c-src containing Gly-63, Arg-95, and Thr-96 with a corresponding fragment of v-src containing Asp-63, Trp-95, and Ile-96 converted p60c-src into a transforming protein by the criteria of focus formation, anchorage-independent growth, and tumor formation in newborn chickens. These mutations also resulted in elevation of the protein kinase activity of p60c-src.

scholarly journals Discovering the first tyrosine kinase

2015 ◽  
Vol 112 (26) ◽  
pp. 7877-7882 ◽  
Author(s):  
Tony Hunter
Keyword(s):  
Tyrosine Kinase ◽  
Kinase Activity ◽  
Human Cancer ◽  
T Antigen ◽  
Protein Kinase Activity ◽  
Rous Sarcoma ◽  
Tumor Viruses ◽  
Sarcoma Virus ◽  
Malignant State

In the middle of the 20th century, animal tumor viruses were heralded as possible models for understanding human cancer. By the mid-1970s, the molecular basis by which tumor viruses transform cells into a malignant state was beginning to emerge as the first viral genomic sequences were reported and the proteins encoded by their transforming genes were identified and characterized. This was a time of great excitement and rapid progress. In 1978, prompted by the discovery from Ray Erikson’s group that the Rous sarcoma virus (RSV) v-Src–transforming protein had an associated protein kinase activity specific for threonine, my group at the Salk Institute set out to determine whether the polyomavirus middle T-transforming protein had a similar kinase activity. Here, I describe the experiments that led to the identification of a kinase activity associated with middle T antigen and our serendipitous discovery that this activity was specific for tyrosine in vitro, and how this in turn led to the fortuitous observation that the v-Src–associated kinase activity was also specific for tyrosine. Our finding that v-Src increased the level of phosphotyrosine in cellular proteins in RSV-transformed cells confirmed that v-Src is a tyrosine kinase and transforms cells by phosphorylating proteins on tyrosine. My colleague Bart Sefton and I reported these findings in the March issue of PNAS in 1980. Remarkably, all of the experiments in this paper were accomplished in less than one month.

scholarly journals Human c-ros-1 gene homologous to the v-ros sequence of UR2 sarcoma virus encodes for a transmembrane receptorlike molecule.

1986 ◽  
Vol 6 (8) ◽  
pp. 3000-3004 ◽  
Author(s):  
H Matsushime ◽  
L H Wang ◽  
M Shibuya
Keyword(s):  
Tyrosine Kinase ◽  
Insulin Receptor ◽  
Transmembrane Domain ◽  
Human Gene ◽  
Kinase Domain ◽  
Glycosylation Site ◽  
Tyrosine Kinase Domain ◽  
Base Pairs ◽  
Human Insulin Receptor ◽  
Sarcoma Virus

We isolated a human gene (designated c-ros-1) homologous to the v-ros sequence of UR2 sarcoma virus. Ten exons, 1,414 base pairs spanning 26 kilobases, contained a tyrosine kinase domain, a transmembrane domain, and a part of an extracellular domain carrying an N glycosylation site which was not acquired by UR2 sarcoma virus. The predicted structure of c-ros-1 is unique among the src family and clearly distinct from the human insulin receptor.

scholarly journals Amino acid substitutions sufficient to convert the nontransforming p60c-src protein to a transforming protein.

1986 ◽  
Vol 6 (12) ◽  
pp. 4155-4160 ◽  
Author(s):  
J Y Kato ◽  
T Takeya ◽  
C Grandori ◽  
H Iba ◽  
J B Levy ◽  
...  
Keyword(s):  
Amino Acid ◽  
Biological Activities ◽  
Tumor Formation ◽  
Protein Kinase Activity ◽  
Focus Formation ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Cellular Homolog ◽  
Sarcoma Virus ◽  
Hybrid Genes

We have previously shown that Rous sarcoma virus variants that carry the cellular homolog (c-src) of the viral src gene (v-src) do not transform chicken embryo fibroblasts. We also have shown that replacement of sequences upstream or downstream from the BglI site of the cellular src gene with the corresponding regions of v-src restored transforming activity to the hybrid genes. Since there are only six amino acid changes between p60c-src and p60v-src within the sequences upstream from BglI, we constructed chimeric molecules involving v-src and c-src to determine the effect of each amino acid substitution on the biological activities of the gene product. We found that the change from Thr to Ile at position 338 or the replacement of a fragment of c-src containing Gly-63, Arg-95, and Thr-96 with a corresponding fragment of v-src containing Asp-63, Trp-95, and Ile-96 converted p60c-src into a transforming protein by the criteria of focus formation, anchorage-independent growth, and tumor formation in newborn chickens. These mutations also resulted in elevation of the protein kinase activity of p60c-src.

scholarly journals Erythropoietin induces tyrosine phosphorylation and kinase activity of the c-fps/fes proto-oncogene product in human erythropoietin-responsive cells

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3193-3196 ◽  
Author(s):  
Y Hanazono ◽  
S Chiba ◽  
K Sasaki ◽  
H Mano ◽  
Y Yazaki ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Tyrosine Phosphorylation ◽  
Kinase Activity ◽  
Protein S ◽  
Receptor Activation ◽  
Kinase Domain ◽  
Cellular Protein ◽  
Erythroid Progenitor ◽  
Nonreceptor Tyrosine Kinase ◽  
Human Erythropoietin

Erythropoietin (EPO) is a hematopoietic growth factor that stimulates the proliferation and differentiation of erythroid progenitor cells. Although the EPO receptor has no kinase domain, EPO rapidly induces tyrosine phosphorylation of several proteins in EPO-responsive cells. Therefore, the receptor activation by the ligand could induce tyrosine- kinase activity of unidentified cellular protein(s). Here we show that c-fps/fes proto-oncogene product (p92c-fes), nonreceptor tyrosine kinase, is tyrosine-phosphorylated on treatment with EPO in a human erythroleukemia cell line TF-1 that is responsive to granulocyte- macrophage colony-stimulating factor, interleukin-3, and EPO. In addition, the kinase activity of p92c-fes was shown to be enhanced by treatment with EPO. Therefore, p92c-fes could be implicated in a signaling pathway triggered by EPO in human EPO-responsive cells.

Lack of induction of neuroretinal cell proliferation by Rous sarcoma virus variants that carry the c-src gene

1985 ◽  
Vol 5 (10) ◽  
pp. 2856-2859
Author(s):  
H Iba ◽  
R Jove ◽  
H Hanafusa
Keyword(s):  
Cell Proliferation ◽  
Rous Sarcoma Virus ◽  
Kinase Activity ◽  
Cell Populations ◽  
Protein Kinase Activity ◽  
Morphological Transformation ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Cellular Homolog ◽  
Sarcoma Virus

Expression of p60v-src of Rous sarcoma virus in cultured chicken embryo neuroretinal cells was previously shown to result in the transformation and sustained proliferation of normally quiescent cell populations. We show here that Rous sarcoma virus variants that encode p60c-src, the cellular homolog of p60v-src, lack the ability to induce morphological transformation and cell proliferation of cultured neuroretinal cells. Neuroretinal cells infected with c-src-containing viruses, however, possess no less p60 protein kinase activity assayed in the immune complex than those infected with the transformation-defective Rous sarcoma virus mutants PA101 or PA104, which do stimulate the growth of these cells.

scholarly journals Lack of induction of neuroretinal cell proliferation by Rous sarcoma virus variants that carry the c-src gene.

1985 ◽  
Vol 5 (10) ◽  
pp. 2856-2859 ◽  
Author(s):  
H Iba ◽  
R Jove ◽  
H Hanafusa
Keyword(s):  
Cell Proliferation ◽  
Rous Sarcoma Virus ◽  
Kinase Activity ◽  
Cell Populations ◽  
Protein Kinase Activity ◽  
Morphological Transformation ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Cellular Homolog ◽  
Sarcoma Virus

Expression of p60v-src of Rous sarcoma virus in cultured chicken embryo neuroretinal cells was previously shown to result in the transformation and sustained proliferation of normally quiescent cell populations. We show here that Rous sarcoma virus variants that encode p60c-src, the cellular homolog of p60v-src, lack the ability to induce morphological transformation and cell proliferation of cultured neuroretinal cells. Neuroretinal cells infected with c-src-containing viruses, however, possess no less p60 protein kinase activity assayed in the immune complex than those infected with the transformation-defective Rous sarcoma virus mutants PA101 or PA104, which do stimulate the growth of these cells.

scholarly journals The FGF Receptor–1 Tyrosine Kinase Domain Regulates Myogenesis but Is Not Sufficient to Stimulate Proliferation

1998 ◽  
Vol 142 (1) ◽  
pp. 241-250 ◽  
Author(s):  
Arthur J. Kudla ◽  
Nathan C. Jones ◽  
R. Scott Rosenthal ◽  
Kirstin Arthur ◽  
Kari L. Clase ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Tyrosine Kinase ◽  
Muscle Cells ◽  
Muscle Differentiation ◽  
Kinase Domain ◽  
Skeletal Muscle Cells ◽  
Tyrosine Kinase Domain ◽  
Protein Kinase Activity ◽  
Skeletal Muscle Differentiation ◽  
Chimeric Receptors

Ligand-stimulated activation of FGF receptors (FGFRs) in skeletal muscle cells represses terminal myogenic differentiation. Skeletal muscle cell lines and subsets of primary cells are dependent on FGFs to repress myogenesis and maintain growth. To understand the intracellular events that transduce these signals, MM14 skeletal muscle cells were transfected with expression vectors encoding chimeric receptors. The chimeras are comprised of the PDGF β receptor (PDGFβR) extracellular domain, the FGFR-1 intracellular domain, and either the PDGFβR or FGFR-1 transmembrane domain. The chimeric receptors were autophosphorylated upon PDGF-BB stimulation and are capable of stimulating mitogen-activated protein kinase activity. Activation of the tyrosine kinase domain of either chimera repressed myogenesis, suggesting intracellular responses regulating skeletal muscle differentiation are transduced by activation of the FGFR-1 tyrosine kinase. Unexpectedly, we found that activation of either chimeric receptor failed to stimulate cellular proliferation. Thus, it appears that regulation of skeletal muscle differentiation by FGFs requires only activation of the FGFR tyrosine kinase. In contrast, stimulation of proliferation may require additional, as yet unidentified, signals involving the receptor ectodomain, the FGF ligand, and heparan sulfate either alone, or in combination.

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