Mitotic cytosol inhibits invagination of coated pits in broken mitotic cells

Marc Pypaert, Dorothy Mundy, Ewen Souter, Jean Claude Labbé, Graham Warren

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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.

Original languageEnglish (US)
Pages (from-to)1159-1166
Number of pages8
JournalJournal of Cell Biology
Volume114
Issue number6
DOIs
StatePublished - Jan 1 1991

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ASJC Scopus subject areas

  • Cell Biology

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Pypaert, M., Mundy, D., Souter, E., Labbé, J. C., & Warren, G. (1991). Mitotic cytosol inhibits invagination of coated pits in broken mitotic cells. Journal of Cell Biology, 114(6), 1159-1166. https://doi.org/10.1083/jcb.114.6.1159