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PMID: 1378446 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Association of p60c-src with endosomal membranes in mammalian fibroblasts.

The Journal of cell biology ·Vol. 118 ·No. 2 ·1992-07-00 ·Pages 321-33

Kaplan KB, Swedlow JR, Varmus HE, Morgan DO

Abstract

We have examined the subcellular localization of p60c-src in mammalian fibroblasts. Analysis of indirect immunofluorescence by three-dimensional optical sectioning microscopy revealed a granular cytoplasmic staining that co-localized with the microtubule organizing center. Immunofluorescence experiments with antibodies against a number of membrane markers demonstrated a striking co-localization between p60c-src and the cation-dependent mannose-6-phosphate receptor (CI-MPR), a marker that identifies endosomes. Both p60c-src and the CI-MPR were found to cluster at the spindle poles throughout mitosis. In addition, treatment of interphase and mitotic cells with brefeldin A resulted in a clustering of p60c-src and CI-MPR at a peri-centriolar position. Biochemical fractionation of cellular membranes showed that a major proportion of p60c-src co-enriched with endocytic membranes. Treatment of membranes containing HRP to alter their apparent density also altered the density of p60c-src-containing membranes. Similar density shift experiments with total cellular membranes revealed that the majority of membrane-associated p60c-src in the cell is associated with endosomes, while very little is associated with plasma membranes. These results support a role for p60c-src in the regulation of endosomal membranes and protein trafficking.

MeSH Terms
Animals Cell Line, Transformed Cytoplasmic Granules/metabolism,ultrastructure Endocytosis Fibroblasts/cytology,metabolism,ultrastructure Fluorescent Antibody Technique Intracellular Membranes/metabolism,ultrastructure Microtubules/metabolism,ultrastructure Mitosis Proto-Oncogene Proteins pp60(c-src)/analysis,genetics,metabolism Rats Transfection
Chemicals
Proto-Oncogene Proteins pp60(c-src)
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kaplan K B
Department of Microbiology, University of California, San Francisco 94143.
Swedlow J R
Varmus H E
Morgan D O
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47 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.118.2.321
Published
1992-07-00
Pages
321-33
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2290043
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.118.2.321 PMC Full Text View journal details
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