ESCRT-III drives the final stages of CUPS maturation for unconventional protein secretion

Abstract

The unconventional secretory pathway exports proteins that bypass the endoplasmic reticulum. In Saccharomyces cerevisiae, conditions that trigger Acb1 secretion via this pathway generate a Grh1 containing compartment composed of vesicles and tubules surrounded by a cup-shaped membrane and collectively called CUPS. Here we report a quantitative assay for Acb1 secretion that reveals requirements for ESCRT-I, -II, and -III but, surprisingly, without the involvement of the Vps4 AAA-ATPase. The major ESCRT-III subunit Snf7 localizes transiently to CUPS and this was accelerated in vps4Δ cells, correlating with increased Acb1 secretion. Microscopic analysis suggests that, instead of forming intraluminal vesicles with the help of Vps4, ESCRT-III/Snf7 promotes direct engulfment of preexisting Grh1 containing vesicles and tubules into a saccule to generate a mature Acb1 containing compartment. This novel multivesicular / multilamellar compartment, we suggest represents the stable secretory form of CUPS that is competent for the release of Acb1 to cells exterior.

DOI: http://dx.doi.org/10.7554/eLife.16299.001

eLife digest

Cells produce thousands of different proteins with a variety of different roles in the body. Some proteins, for example the hormone insulin, perform roles outside of the cell and are released from cells in a process that has several stages. In the first step, newly-made insulin and many other “secretory” proteins enter a compartment called the endoplasmic reticulum. Once inside, these proteins can then be loaded into other compartments and transported to the edge of the cell.

There is another class of secretory proteins that are released from the cell without first entering the endoplasmic reticulum, in a process termed “unconventional protein secretion”. A protein called Acb1 is released from yeast cells in this manner. Previous research identified a compartment that might be involved in this process. However, it is not clear how this compartment (named CUPS) forms, and what role it plays in unconventional protein secretion.

Curwin et al. investigated how CUPS form in yeast cells, and whether the compartment contains Acb1 proteins. The experiments reveal that after CUPS form they need to mature into a form that is involved in the release of Acb1 proteins from the cell. This maturation process involves some, but not all, of the same genes as those involved in producing another type of compartment in cells called a multivesicular body. Acb1 is only found in the mature CUPS and multivesicular bodies are not involved in the release of this protein from the cell.

Curwin et al.’s findings shed some light on how Acb1 and other secretory proteins can be released from cells without involving the endoplasmic reticulum. Future challenges are to reveal how CUPS capture cargo and find out how Acb1 leaves the CUPS to exit the cell.

DOI: http://dx.doi.org/10.7554/eLife.16299.002