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PMID: 1910051 Published · ppublish English Journal Article

Mitotic cytosol inhibits invagination of coated pits in broken mitotic cells.

The Journal of cell biology ·Vol. 114 ·No. 6 ·1991-09-00 ·Pages 1159-66

Pypaert M, Mundy D, Souter E, Labbé JC, Warren G

Abstract

Receptor-mediated endocytosis is inhibited during mitosis in mammalian cells and earlier work on A431 cells suggested that one of the sites inhibited was the invagination of coated pits (Pypaert, M., J. M. Lucocq, and G. Warren. 1987. Eur. J. Cell Biol. 45: 23-29). To explore this inhibition further, we have reproduced it in broken HeLa cells. Mitotic or interphase cells were broken by freeze-thawing in liquid nitrogen and warmed in the presence of mitotic or interphase cytosol. Using a morphological assay, we found invagination to be inhibited only when mitotic cells were incubated in mitotic cytosol. This inhibition was reversed by diluting the cytosol during the incubation. Reversal was sensitive to okadaic acid, a potent phosphatase inhibitor, showing that phosphorylation was involved in the inhibition of invagination. This was confirmed using purified cdc2 kinase which alone could partially substitute for mitotic cytosol.

MeSH Terms
CDC2 Protein Kinase/isolation & purification,metabolism Coated Pits, Cell-Membrane/drug effects,physiology,ultrastructure Cytosol/physiology Endocytosis Ethers, Cyclic/pharmacology HeLa Cells/cytology,physiology Humans Interphase/physiology Microscopy, Electron Mitosis/drug effects,physiology Okadaic Acid
Chemicals
Ethers, Cyclic Okadaic Acid CDC2 Protein Kinase
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Pypaert M
Imperial Cancer Research Fund, London, England.
Mundy D
Souter E
Labbé J C
Warren G
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41 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.114.6.1159
Published
1991-09-00
Pages
1159-66
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2289130
Subset
IM
External Links
PubMed source DOI: 10.1083/jcb.114.6.1159 PMC Full Text View journal details
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