Immunofluorescence localization of phosphotyrosine containing proteins in RSV-transformed mouse fibroblasts

Research output: Contribution to journalArticle

Abstract

The localization of phosphotyrosine-containing proteins (P-TYR proteins) was studied by immunofluorescence microscopy employing affinity-purified azobenzyl phosphonate (ABP) antibodies, which specifically cross-react with phosphotyrosine. In Rous sarcoma virus (RSV)-transformed fibroblasts, after fixation followed by permeabilization with nonionic detergents, ABP antibodies gave a diffuse staining of the cytoplasm and specifically decorated restricted areas of the ventral plasma membrane corresponding to adhesion plaques, identified by interference reflection microscopy and staining with anti-vinculin serum. Specific decoration was also observed at the level of cell-cell contacts and at the tips of filopodial protrusions. Control non-transformed 3T3 mouse fibroblasts were not significantly stained by ABP antibodies. These findings show that, in RSV-transformed cells, proteins phosphorylated at tyrosine residues are found at cell-substratum and cell-cell contacts.

Original languageEnglish
Pages (from-to)112-124
Number of pages13
JournalExperimental Cell Research
Volume154
Issue number1
DOIs
Publication statusPublished - 1984

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Rous sarcoma virus
Phosphotyrosine
Fluorescent Antibody Technique
Fibroblasts
Organophosphonates
Proteins
Antibodies
Interference Microscopy
Staining and Labeling
Vinculin
Focal Adhesions
Fluorescence Microscopy
Detergents
Tyrosine
Cytoplasm
Cell Membrane
Serum

ASJC Scopus subject areas

  • Cell Biology

Cite this

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title = "Immunofluorescence localization of phosphotyrosine containing proteins in RSV-transformed mouse fibroblasts",
abstract = "The localization of phosphotyrosine-containing proteins (P-TYR proteins) was studied by immunofluorescence microscopy employing affinity-purified azobenzyl phosphonate (ABP) antibodies, which specifically cross-react with phosphotyrosine. In Rous sarcoma virus (RSV)-transformed fibroblasts, after fixation followed by permeabilization with nonionic detergents, ABP antibodies gave a diffuse staining of the cytoplasm and specifically decorated restricted areas of the ventral plasma membrane corresponding to adhesion plaques, identified by interference reflection microscopy and staining with anti-vinculin serum. Specific decoration was also observed at the level of cell-cell contacts and at the tips of filopodial protrusions. Control non-transformed 3T3 mouse fibroblasts were not significantly stained by ABP antibodies. These findings show that, in RSV-transformed cells, proteins phosphorylated at tyrosine residues are found at cell-substratum and cell-cell contacts.",
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AU - Di Renzo, Maria Flavia

AU - Comoglio, Paolo M.

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