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Identification and Localization of Phosphoproteins in v-onc Transformed Fibroblasts by Means of Phosphotyrosine Antibodies

  • P. M. Comoglio
  • D. Cirillo
  • M. F. Di Renzo
  • R. Ferracini
  • F. G. Giancotti
  • S. Giordano
  • L. Naldini
  • G. Tarone
  • P. C. Marchisio
Part of the NATO ASI Series book series (NSSA, volume 91)

Abstract

The transformation process induced by several retroviruses, includ­ing Rous sarcoma virus (RSV), Feline sarcoma virus (FeSV), Fujinami avian sarcoma virus (FSV) and Abelson murine leukemia virus (AMuLV), is triggered and maintained by the action of v-onc genes which all code for transforming proteins with associated tyrosine kinase activity (for review see 1). Since protein phosphorylation seems to be invariably associated with molecular mechanism(s) involved in growth control and in the neoplastic transformation triggered by these retroviruses, the identification of cellular proteins phosphorylated at tyrosine residues is an issue of major importance. Putative substrates of tyrosine kinases have been identified by means of conventional techniques such as bidimensional separation of total cellular proteins followed by phosphoaminoacid analysis. However, these techniques seem to have intrinsic limitations - as shown also by the failure to identify well known substrates - such as the transforming proteins themselves, which are known to be heavily tyrosine-phosphorylated. The difficulties are generated by the fact that phosphotyrosine represents less than 2% of phospho­aminoacids also in transformed cells (being less than 0.2% in nor­mal cells); in addition it has been shown that, only a minor frac­tion, i.e. less than 10%, of each substrate molecules of v-onc coded kinases, is phosphorylated at tyrosine even in fully trans­formed cells.

Keywords

Rous Sarcoma Virus Phenyl Phosphate Adhesion Plaque Transforming Protein Detergent Insoluble Fraction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • P. M. Comoglio
    • 1
  • D. Cirillo
    • 1
  • M. F. Di Renzo
    • 1
  • R. Ferracini
    • 1
  • F. G. Giancotti
    • 1
  • S. Giordano
    • 1
  • L. Naldini
    • 1
  • G. Tarone
    • 1
  • P. C. Marchisio
    • 1
  1. 1.Institute of HistologyUniversity of Torino Medical SchoolTorinoItaly

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