Membrane Budding  James H. Hurley, Evzen Boura, Lars-Anders Carlson, Bartosz Różycki  Cell  Volume 143, Issue 6, Pages 875-887 (December 2010) DOI: 10.1016/j.cell.2010.11.030 Copyright © 2010 Elsevier Inc. Terms and Conditions

Figure 1 Proteins and Lipid Microdomains in Membrane Budding (A) Budding of phase-separated lipid microdomains from GUVs (giant unilamellar vesicles) composed of synthetic lipids is an example of membrane budding in the absence of any proteins. Reproduced by permission from Baumgart et al. (2003). (B) Shiga toxin (black dots) acts from outside the plasma membrane to induce membrane buds and is an example of a protein triggering budding events that are primarily driven by lipid microdomains. Image reproduced by permission from Macmillan Publishers Ltd: Nature, Römer et al. (2007), copyright 2007. (C) Budding by caveolae represents a hybrid between a membrane microdomain and protein coat-driven mechanisms. Reproduced by permission from Macmillan Publishers Ltd: Nat. Rev. Mol. Cell. Biol., Parton and Simons (2007), copyright 2007. (D) ESCRT-I and -II induce buds in synthetic GUVs. Reproduced by permission from Wollert and Hurley (2010). Proteins organize these structures but do not form a coat, suggesting a possible role for microdomains. (E) HIV-1 buds visualized by electron tomography (Carlson et al., 2008). The bud is organized by the HIV-1 capsid protein, heavily enriched in raft lipids, and cleaved by ESCRT proteins. (F) Deep etch visualization of clathrin-coated pits (image courtesy of J. Heuser). Clathrin assembles into baskets in the absence of membranes but is thought to be too flexible to deform membranes on its own. For this, clathrin needs help from other membrane-deforming proteins and possibly from lipids. (G) The COP II cage is an example of a protein structure that can form in the absence of lipids and can impose its shape on any simple bilayer-forming lipid mixture. Reproduced by permission from Russell and Stagg (2010). Cell 2010 143, 875-887DOI: (10.1016/j.cell.2010.11.030) Copyright © 2010 Elsevier Inc. Terms and Conditions

Figure 2 Coated Vesicle Budding (A) Structure of a clathrin basket from cytoelectron microscopy; reproduced by permission from Macmillan Publishers Ltd: Nature, Fotin et al. (2004), copyright 2004. (B) COP II vesicles produced from purified components; reproduced by permission from Lee et al. (2005). (C) Structural parallels between clathrin, COP I, and COP II. Adapted from Lee and Goldberg (2010). Cell 2010 143, 875-887DOI: (10.1016/j.cell.2010.11.030) Copyright © 2010 Elsevier Inc. Terms and Conditions

Figure 3 Membrane Microdomains and Budding (A) Coexistence of phases in model membranes visualized by atomic force microscopy in a supported bilayer (a membrane bilayer adsorbed onto a solid support, usually glass). Reproduced with permission from Chiantia et al. (2006). (B) Phase transitions in a single-lipid membrane analyzed by molecular dynamics simulations. Reproduced with permission from Heller et al. (1993). Copyright 1993 American Chemical Society. (C) Schematic model of a raft-type membrane microdomain, including a model of a myristoylated ESCRT-III subunit Vps20 as an example of protein that might anchor to rafts. Cell 2010 143, 875-887DOI: (10.1016/j.cell.2010.11.030) Copyright © 2010 Elsevier Inc. Terms and Conditions

Figure 4 Protein Structures that Cluster Raft Lipids (A) Simian virus 40 VP1 pentamer bound to the membrane via the headgroup of the ganglioside GM1 (Neu et al., 2008). (B) Cholera toxin B subunit pentamer bound to GM1 (Merritt et al., 1994). (C) Composite model of the myristoylated HIV-1 matrix domain trimer bound to PI(4,5)P2 (Hill et al., 1996; Saad et al., 2006, 2008). In each case, lipid tails are modeled. Images were generated with VMD 1.8.6. Cell 2010 143, 875-887DOI: (10.1016/j.cell.2010.11.030) Copyright © 2010 Elsevier Inc. Terms and Conditions

Figure 5 Multivesicular Bodies Bud via Diverse Mechanisms (A) Multivesicular bodies (MVBs) form from late endosomes in animal cells. Their formation is dependent on both ESCRT complexes and the unusual lipid lysobisphosphatidic acid (LBPA). (B) The conserved ESCRT-dependent MVB biogenesis pathway from early endosomes in yeast and animal cells. PI(3)P has been directly visualized in these MVBs. Cholesterol has been visualized in MVBs from animal cells, but it has not been directly confirmed whether these are ESCRT dependent or not. (C) Specialized formation of MVBs containing polymerized Pmel17. (D) Ceramide-dependent MVBs bud from raft-like and tetraspanin-enriched microdomains in animal cells. Cell 2010 143, 875-887DOI: (10.1016/j.cell.2010.11.030) Copyright © 2010 Elsevier Inc. Terms and Conditions

Figure 6 Lipids and ESCRTs in HIV-1 Assembly Apart from viral proteins, the release of HIV-1 requires both specific cellular lipids and proteins, which are recruited to the budding site by the viral Gag protein. Gag assembles into an imperfect hexagonal lattice on the plasma membrane (Briggs et al., 2009). It binds the plasma membrane marker PI(4,5)P2 through a specific binding site in its N terminus. PI(4,5)P2, cholesterol, and certain other raft lipids are enriched in the viral membrane compared to the plasma membrane. Through its C terminus, Gag recruits the ESCRT proteins to the budding site. Gag can bind both ESCRT-I and ALIX, which both recruit ESCRT-III to the budding site. Cell 2010 143, 875-887DOI: (10.1016/j.cell.2010.11.030) Copyright © 2010 Elsevier Inc. Terms and Conditions