scholarly journals Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex.

1983 ◽  
Vol 3 (1) ◽  
pp. 9-19 ◽  
Author(s):  
J Brugge ◽  
W Yonemoto ◽  
D Darrow
Keyword(s):  
Rous Sarcoma Virus ◽  
Temperature Sensitive ◽  
Chase Period ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Discrete Population ◽  
Sarcoma Virus ◽  
Transforming Activity ◽  
The Stability ◽  
In Cells

The transforming protein of Rous sarcoma virus (RSV), pp60src, was previously shown to associate with two cellular proteins of Mr 90,000 and 50,000 in RSV-transformed chicken cells. In this report, we demonstrate that this interaction is specific for a discrete population of pp60src molecules. Newly synthesized pp60src was found to preferentially associate with pp90 and pp50 to form a short-lived complex. The half-life of this complex varied from 9 to 15 min in cells transformed by nondefective strains of RSV. This interaction between pp60src, pp50, and pp90 took place in a soluble fraction of the cell, and the complex-bound pp60src molecules were not phosphorylated on tyrosine. These results suggest that pp90 and pp50 may be involved in the processing of pp60src molecules before the association of pp60src with the plasma membrane. The kinetics of dissociation of this complex were shown to be altered in cells infected with viruses containing a temperature-sensitive defect in the src gene. When cells infected with these viruses were grown at the nonpermissive temperature, more than 90% of the pp60src molecules were associated with pp90 and pp50, and little or no dissociation was observed in a 3-h chase period. These results suggest that mutations in the src gene which affect the transforming activity of pp60src also affect the stability of the interaction of pp60src with pp90 and pp50.

Interaction between the Rous sarcoma virus transforming protein and two cellular phosphoproteins: analysis of the turnover and distribution of this complex

1983 ◽  
Vol 3 (1) ◽  
pp. 9-19
Author(s):  
J Brugge ◽  
W Yonemoto ◽  
D Darrow
Keyword(s):  
Rous Sarcoma Virus ◽  
Temperature Sensitive ◽  
Chase Period ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Discrete Population ◽  
Sarcoma Virus ◽  
Transforming Activity ◽  
The Stability ◽  
In Cells

The transforming protein of Rous sarcoma virus (RSV), pp60src, was previously shown to associate with two cellular proteins of Mr 90,000 and 50,000 in RSV-transformed chicken cells. In this report, we demonstrate that this interaction is specific for a discrete population of pp60src molecules. Newly synthesized pp60src was found to preferentially associate with pp90 and pp50 to form a short-lived complex. The half-life of this complex varied from 9 to 15 min in cells transformed by nondefective strains of RSV. This interaction between pp60src, pp50, and pp90 took place in a soluble fraction of the cell, and the complex-bound pp60src molecules were not phosphorylated on tyrosine. These results suggest that pp90 and pp50 may be involved in the processing of pp60src molecules before the association of pp60src with the plasma membrane. The kinetics of dissociation of this complex were shown to be altered in cells infected with viruses containing a temperature-sensitive defect in the src gene. When cells infected with these viruses were grown at the nonpermissive temperature, more than 90% of the pp60src molecules were associated with pp90 and pp50, and little or no dissociation was observed in a 3-h chase period. These results suggest that mutations in the src gene which affect the transforming activity of pp60src also affect the stability of the interaction of pp60src with pp90 and pp50.

Functional domains of the pp60v-src protein as revealed by analysis of temperature-sensitive Rous sarcoma virus mutants

1984 ◽  
Vol 4 (8) ◽  
pp. 1508-1514
Author(s):  
A W Stoker ◽  
P J Enrietto ◽  
J A Wyke
Keyword(s):  
Rous Sarcoma Virus ◽  
Kinase Activity ◽  
Temperature Sensitive ◽  
Tyrosine Kinase Activity ◽  
Rous Sarcoma ◽  
Heat Labile ◽  
Src Gene ◽  
Sarcoma Virus

Four temperature-sensitive (ts) Rous sarcoma virus src gene mutants with lesions in different parts of the gene represent three classes of alteration in pp60src. These classes are composed of mutants with (i) heat-labile protein kinase activities both in vitro and in vivo (tsLA27 and tsLA29), (ii) heat-labile kinases in vivo but not in vitro (tsLA33), and (iii) neither in vivo nor in vitro heat-labile kinases (tsLA32). The latter class indicates the existence of structural or functional pp60src domains that are required for transformation but do not grossly affect tyrosine kinase activity.

scholarly journals A glycoprotein in the plasma membrane matrix as a major potential substrate of p60v-src.

1990 ◽  
Vol 10 (2) ◽  
pp. 830-836 ◽  
Author(s):  
M Hamaguchi ◽  
M Matsuda ◽  
H Hanafusa
Keyword(s):  
Plasma Membrane ◽  
Rous Sarcoma Virus ◽  
Transmembrane Protein ◽  
Temperature Sensitive ◽  
Morphological Transformation ◽  
Fibronectin Receptor ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Potential Substrate ◽  
In Cells

A potential substrate of p60v-src in Rous sarcoma virus-transformed cells was found to be a 130-kilodalton (kDa) glycoprotein which binds to lectin-Sepharose and can be immunoprecipitated by an anti-phosphotyrosine antibody. This glycoprotein was shown to be distinct from the fibronectin receptor and a cellular protein phosphorylated in p60v-src immune complexes. The protein was a transmembrane protein localized in the plasma membrane and resistant to extraction with Triton X-100. The 130-kDa protein was also highly phosphorylated in cells transformed by Fujinami sarcoma virus or Y73 but not in cells infected with Rous sarcoma virus mutants that encode p60v-src lacking myristoylated N termini. Phosphorylation of this glycoprotein was temperature dependent in cells infected with temperature-sensitive mutants. The good correlation between its phosphorylation and morphological transformation, together with its relative abundance among phosphorylated proteins and its subcellular localization, suggests that phosphorylation of the 130-kDa glycoprotein is one of the primary events important for cell transformation by p60v-src and related oncogene products.

Phosphorylation of talin at tyrosine in Rous sarcoma virus-transformed cells

1987 ◽  
Vol 7 (1) ◽  
pp. 371-378
Author(s):  
J E DeClue ◽  
G S Martin
Keyword(s):  
Rous Sarcoma Virus ◽  
Peptide Mapping ◽  
Temperature Sensitive ◽  
Permissive Temperature ◽  
Sensitive Mutant ◽  
Tyrosine Residues ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Chicken Embryo Fibroblasts ◽  
In Cells

The cytoskeletal protein talin was found to undergo enhanced phosphorylation at tyrosine residues in chicken embryo fibroblasts following transformation by Rous sarcoma virus. An increase in the tyrosine phosphorylation of talin was also observed within 6 h in cells infected by the temperature-sensitive mutant tsNY68 after a shift from the nonpermissive to the permissive temperature. The overall extent of phosphorylation was 0.07 mol of phosphate per mol of talin and was not appreciably altered by transformation. In uninfected cells talin was shown to be phosphorylated at multiple sites by tryptic peptide mapping. Following transformation most of these sites remained phosphorylated, to the same or to a lesser extent, while novel, phosphotyrosine-containing phosphopeptides appeared. Talin was phosphorylated at tyrosine in cells infected by Rous sarcoma virus mutants which induce altered or partial transformation morphologies; thus the increased phosphorylation of talin at tyrosine occurred irrespective of the morphology induced. Transformation by Y73 also induced elevated levels of phosphotyrosine in talin, whereas transformation by the avian erythroblastosis and Fujinami sarcoma viruses did not.

A glycoprotein in the plasma membrane matrix as a major potential substrate of p60v-src

1990 ◽  
Vol 10 (2) ◽  
pp. 830-836
Author(s):  
M Hamaguchi ◽  
M Matsuda ◽  
H Hanafusa
Keyword(s):  
Plasma Membrane ◽  
Rous Sarcoma Virus ◽  
Transmembrane Protein ◽  
Temperature Sensitive ◽  
Morphological Transformation ◽  
Fibronectin Receptor ◽  
Rous Sarcoma ◽  
Sarcoma Virus ◽  
Potential Substrate ◽  
In Cells

A potential substrate of p60v-src in Rous sarcoma virus-transformed cells was found to be a 130-kilodalton (kDa) glycoprotein which binds to lectin-Sepharose and can be immunoprecipitated by an anti-phosphotyrosine antibody. This glycoprotein was shown to be distinct from the fibronectin receptor and a cellular protein phosphorylated in p60v-src immune complexes. The protein was a transmembrane protein localized in the plasma membrane and resistant to extraction with Triton X-100. The 130-kDa protein was also highly phosphorylated in cells transformed by Fujinami sarcoma virus or Y73 but not in cells infected with Rous sarcoma virus mutants that encode p60v-src lacking myristoylated N termini. Phosphorylation of this glycoprotein was temperature dependent in cells infected with temperature-sensitive mutants. The good correlation between its phosphorylation and morphological transformation, together with its relative abundance among phosphorylated proteins and its subcellular localization, suggests that phosphorylation of the 130-kDa glycoprotein is one of the primary events important for cell transformation by p60v-src and related oncogene products.

Transformation-defective, Temperature-sensitive Mutants of Rous Sarcoma Virus Have a Reversibly Defective src-gene Product

1980 ◽  
Vol 44 (0) ◽  
pp. 1007-1012 ◽  
Author(s):  
R. R. Friis ◽  
B. M. Jockusch ◽  
C. B. Boschek ◽  
A. Ziemiecki ◽  
H. Rubsamen ◽  
...  
Keyword(s):  
Gene Product ◽  
Rous Sarcoma Virus ◽  
Temperature Sensitive ◽  
Temperature Sensitive Mutants ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Sarcoma Virus

scholarly journals N-terminal deletions in Rous sarcoma virus p60src: effects on tyrosine kinase and biological activities and on recombination in tissue culture with the cellular src gene.

1985 ◽  
Vol 5 (10) ◽  
pp. 2789-2795 ◽  
Author(s):  
F R Cross ◽  
E A Garber ◽  
H Hanafusa
Keyword(s):  
Amino Acids ◽  
Tyrosine Kinase ◽  
Rous Sarcoma Virus ◽  
Kinase Activity ◽  
Biological Activities ◽  
Tyrosine Kinase Activity ◽  
Rous Sarcoma ◽  
Src Gene ◽  
Sarcoma Virus ◽  
Transforming Activity

We have constructed deletions within the region of cloned Rous sarcoma virus DNA coding for the N-terminal 30 kilodaltons of p60src. Infectious virus was recovered after transfection. Deletions of amino acids 15 to 149, 15 to 169, or 149 to 169 attenuated but did not abolish transforming activity, as assayed by focus formation and anchorage-independent growth. These deletions also had only slight effects on the tyrosine kinase activity of the mutant src protein. Deletion of amino acids 169 to 264 or 15 to 264 completely abolished transforming activity, and src kinase activity was reduced at least 10-fold. However, these mutant viruses generated low levels of transforming virus by recombination with the cellular src gene. The results suggest that as well as previously identified functional domains for p60src myristylation and membrane binding (amino acids 1 to 14) and tyrosine kinase activity (amino acids 250 to 526), additional N-terminal sequences (particularly amino acids 82 to 169) can influence the transforming activity of the src protein.

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