Additional file 1: Additional Material 1-Supplementary Figure 1. The ESCRT machinery is composed of five sub-complexes each functioning consecutively for recruitment of the downstream subcomplex. The process begins with the recruitment of ESCRT0 or its analogue TOM1-esc for recognition of tagged Ubiquitin on cargo, and endosomal membrane phospholipids such as phosphatidylinositol 3-phosphate (PtdIns [3]P) upon which the ESCRTI, composed of VPS23, VPS28, and VPS37, is recruited, with its only known role being ubiquitin recognition via its UIM domain [18]. The assembly of ESCRTI then leads to assembly of the heterotetrameric ESCRTII consisting of VPS36, VPS22, and two copies of VPS25 which also bind to PtdIns [3]P via the FYVE domains [18]. Finally, this leads to the recruitment of the ESCRTIII machinery, a heteropentameric complex consisting of SNF7-domain containing family proteins, VPS20, VPS32, VPS2, VPS24, and CHMP7 [18]. A filamentous VPS32 polypeptide capped by VPS2 and VPS24 (also belonging to the paralagous SNF7-domain containing family of proteins) induces ILV formation by constricting the neck of the budding vesicle, a process which is catalysed by the ESCRTIIIA VPS4, an AAA+ ATPase [18]. It is also hypothesized that ESCRTIIIA components such as VPS31 and VPS46 are required for stabilizing the sub-complexes during the budding processes while others are needed for recycling of the complexes back into the cytosol once the process is complete [18]. Figure adapted from Stenmark and Raiborg [18].