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PMID: 1702788 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Immunolocalization of the cellular src protein in interphase and mitotic NIH c-src overexpresser cells.

The Journal of cell biology ·Vol. 111 ·No. 6 Pt 2 ·1990-12-00 ·Pages 3097-116

David-Pfeuty T, Nouvian-Dooghe Y

Abstract

The mouse mAb, mAb 327, that recognizes specifically both pp60v-src and pp60c-src in a wide variety of cells, has been used to determine precisely the various locations of pp60c-src in NIH c-src overexpresser cells, using the technique of immunofluorescence microscopy. In interphase cells, the protein exhibits two main distributions: one that appears uniform and in association with the cell surface and the other that is patchy and juxtanuclear and coincides with the centrosomes. The juxtanuclear aggregation of pp60c-src-containing patches depends on microtubules and does not seem to occur within the Golgi apparatus and the rough ER. At the G2-to-M-phase transition, a drastic change in the localization patterns of pp60c-src takes place. We also report experiments in which the NIH c-src overexpresser cells were exposed to Con A for various times to induce a redistribution of the cell surface Con A receptors. We show that, at each stage of the Con A-mediated endocytotic process, the Con A-receptor complexes redistribute into structures to which pp60c-src appears also to be associated: at first, into patches that form at the cell surface level and then, into a cap that stands at the cell center in a juxtanuclear position and that coincides with the Golgi apparatus. During this capping process, pp60c-src-containing vesicles continue to accumulate in a centriolar spot, as in interphase, Con A-untreated cells, from which Con A is excluded. The significance of the intracellular locations of pp60c-src to the possible functions of the protein is discussed. Also, the distribution patterns of the cellular protein in the NIH c-src overexpresser cells are compared with those of pp60v-src in RSV-transformed cells. The differences observed are discussed in relation with the differences in transforming capacities of the two proteins. Finally, the possible physiological significance of the association between pp60c-src and the structures generated after the binding of Con A to its surface receptors is addressed.

MeSH Terms
Animals Cell Adhesion/physiology Cell Compartmentation Cell Line Concanavalin A/metabolism Endocytosis/physiology Fluorescent Dyes Immunohistochemistry Interphase/physiology Mitosis/physiology Proto-Oncogene Proteins pp60(c-src)/analysis,physiology Recombinant Proteins/analysis Transfection
Chemicals
Fluorescent Dyes Recombinant Proteins Concanavalin A Proto-Oncogene Proteins pp60(c-src)
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
David-Pfeuty T
Institut Curie-Biologie, Centre Universitaire, Orsay, France.
Nouvian-Dooghe Y
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.111.6.3097
Published
1990-12-00
Pages
3097-116
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2116410
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.111.6.3097 PMC Full Text View journal details
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