Thu, 21 Aug 2008 09:35:19 BST CiteULike: yunpeics clathrin http://www.citeulike.org/user/yunpeic/tag/clathrin CiteULike.org en-gb Copyright © 2004-2008 citeulike.org http://www.citeulike.org/user/yunpeic/article/1697093 <i>Exp Cell Res, Vol. 312, No. 20. (10 December 2006), pp. 4036-4048.</i><br /><br />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 (PIP(2)). 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 PIP(2) 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 approximately 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. Role of lipids and actin in the formation of clathrin-coated pits. E Boucrot S Saffarian R Massol T Kirchhausen M Ehrlich doi:10.1016/j.yexcr.2006.09.025 Exp Cell Res, Vol. 312, No. 20. (10 December 2006), pp. 4036-4048. 2007-09-26T12:14:35-00:00 2006 Exp Cell Res 0014-4827 312 20 4036 4048 clathrin imaging tirf traffic widefield http://www.citeulike.org/user/yunpeic/article/2091284 <i>Nat Cell Biol, Vol. 4, No. 9. (2002), pp. 691-698.</i> Imaging actin and dynamin recruitment during invagination of single clathrin-coated pits Christien Merrifield Morris Feldman Lei Wan Wolfhard Almers doi:10.1038/ncb837 Nat Cell Biol, Vol. 4, No. 9. (2002), pp. 691-698. 2007-12-11T20:04:08-00:00 2002 Nat Cell Biol 4 9 691 698 clathrin imaging single_molecule tirf widefield http://www.citeulike.org/user/yunpeic/article/2008552 <i>Cell, Vol. 118, No. 5. (3 September 2004), pp. 591-605.</i><br /><br />Clathrin-coated vesicles carry traffic from the plasma membrane to endosomes. We report here the real-time visualization of cargo sorting and endocytosis by clathrin-coated pits in living cells. We have detected the formation of coats by monitoring incorporation of fluorescently tagged clathrin or its adaptor AP-2; we have also followed clathrin-mediated uptake of transferrin and of single LDL or reovirus particles. The intensity of a cargo-loaded clathrin cluster grows steadily during its lifetime, and the time required to complete assembly is proportional to the size of the cargo particle. These results are consistent with a nucleation-growth mechanism and an approximately constant growth rate. There are no strongly preferred nucleation sites. A proportion of the nucleation events are weak and short lived. Cargo incorporation occurs primarily or exclusively in a newly formed coated pit. Our data lead to a model in which coated pits initiate randomly but collapse unless stabilized, perhaps by cargo capture. Endocytosis by Random Initiation and Stabilization of Clathrin-Coated Pits Marcelo Ehrlich Werner Boll Antoine van Oijen Ramesh Hariharan Kartik Chandran Max Nibert Tomas Kirchhausen doi:10.1016/j.cell.2004.08.017 Cell, Vol. 118, No. 5. (3 September 2004), pp. 591-605. 2007-11-29T00:02:46-00:00 2004 Cell 118 5 591 605 clathrin endocytosis