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PMID: 6982901 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Epidermal growth factor-induced centrosomal separation: mechanism and relationship to mitogenesis.

The Journal of cell biology ·Vol. 95 ·No. 1 ·1982-10-00 ·Pages 316-22

Sherline P, Mascardo R

Abstract

Using a rabbit antibody to MAP1 to stain centrosomes we have studied the mechanism by which epidermal growth factor (EGF) induces centrosomal separation in HeLa cells. The response is rapid, being detectable within 20 min after EGF (100 ng/ml) addition and by 4 h 40% of logarithmically growing cells and greater than 70% of cells synchronized at G1/S with 1 mM hydroxyurea show centrosomes separated by more than one diameter. A concentration of 0.05 ng/ml of EGF induces significant separation in synchronized cells (5-9% control vs. 20% with EGF at 0.05 ng/ml) and 0.1 to 0.5 ng/ml induces a half maximal response. Centrosomal separation is blocked by energy inhibitors, trifluoperazine, chlorpromazine, and W-7, cytochalasins B and D, and taxol, and is stimulated or enhanced by A23187, colchicine, and oncodazole. Trifluoperazine, W-7, cytochalasin D, and taxol also block DNA synthesis in response to EGF as measured by autoradiography using [3H]thymidine. Our hypothesis based upon these results is that EGF, by raising the free calcium level, activates calmodulin, which stimulates contraction of microfilaments attached to the centrosome, pulling the daughter centrosome apart. EGF may also induce depolymerization or detachment of microtubules in the vicinity of the centrosome which ordinarily serve to maintain its position and inhibit separation. Centrosomal separation may be a key event in triggering DNA synthesis in response to EGF and colchicine.

MeSH Terms
Calcium/physiology Calmodulin/physiology Cell Division/drug effects Centrioles/physiology DNA/biosynthesis Energy Metabolism Epidermal Growth Factor/pharmacology Humans Microtubules/physiology Mitosis/drug effects Organoids/physiology Time Factors
Chemicals
Calmodulin Epidermal Growth Factor DNA Calcium
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Sherline P
Mascardo R
References (52)
52 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.95.1.316
Published
1982-10-00
Pages
316-22
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2112368
Subset
IM
Grants
NIGMS NIH HHS · GM-22497 · United States
External Links
PubMed source DOI: 10.1083/jcb.95.1.316 PMC Full Text View journal details
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