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    Role of lipids and actin in the formation of clathrin-coated pits

    Boucrot, Emmanuel and Saffarian, S. and Massol, R. and Kirchhausen, T. and Ehrlich, M. (2006) Role of lipids and actin in the formation of clathrin-coated pits. Experimental Cell Research 312 (20), pp. 4036-4048. ISSN 0014-4827.

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    Abstract

    Assembly of clathrin-coated pits and their maturation into coated vesicles requires coordinated interactions between specific lipids and several structural and regulatory proteins. In the presence of primary alcohols, phospholipase D generates phosphatidylalcohols instead of PA, reducing stimulation of phosphatidyl inositol 5-kinase (PI5K) and hence decreasing formation of phosphoinositide-4,5-biphosphate (PIP2). Using live-cell imaging, we have shown that acute treatment of cells with 1-butanol or other small primary alcohols induces rapid disassembly of coated pits at the plasma membrane and blocks appearance of new ones. Addition of exogenous PIP2 reverses this effect. Coated pits and vesicles reappear synchronously upon removal of 1-butanol; we have used this synchrony to assess the role of actin in coated vesicle assembly. Prolonged inhibition of actin polymerization by latrunculin A or cytochalasin D reduced by ∼ 50% the frequency of coated pit formation without affecting maturation into coated vesicles. As in control cells, removal of 1-butanol in the continued presence of an actin depolymerizer led to synchronous appearance of new pits, which matured normally. Thus, remodeling of the actin cytoskeleton is not essential for clathrin-coated vesicle assembly but may indirectly affect the nucleation of clathrin-coated pits.

    Metadata

    Item Type: Article
    School: School of Law
    Depositing User: Sarah Hall
    Date Deposited: 07 May 2019 12:07
    Last Modified: 07 May 2019 12:07
    URI: https://eprints.bbk.ac.uk/id/eprint/27421

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