Toxin pores endocytosed during plasma membrane repair traffic into the lumen of MVBs for degradation.
Cells permeabilized by the bacterial pore-forming toxin streptolysin O (SLO) reseal their plasma membrane in a Ca(2+) -dependent manner. Resealing involves Ca(2+) -dependent exocytosis of lysosomes, release of acid sphingomyelinase and rapid formation of endosomes that carry the transmembrane pores into the cell. The intracellular fate of the toxin-carrying endocytic vesicles, however, is still unknown. Here, we show that SLO pores removed from the plasma membrane by endocytosis are sorted into the lumen of lysosomes, where they are degraded. SLO-permeabilized cells contain elevated numbers of total endosomes, which increase gradually in size while transitioning from endosomes with flat clathrin coats to large multivesicular bodies (MVBs). Under conditions that allow endocytosis and plasma membrane repair, SLO is rapidly ubiquitinated and gradually degraded, in a process sensitive to inhibitors of lysosomal hydrolysis but not of proteasomes. The endosomes induced by SLO permeabilization become increasingly acidified and promote SLO degradation under normal conditions, but not in cells silenced for expression of Vps24, an ESCRT-III complex component required for the release of intraluminal vesicles into MVBs. Thus, cells dispose of SLO transmembrane pores by ubiquitination/ESCRT-dependent sorting into the lumen of late endosomes/lysosomes.
Department of Cell Biology and Molecular Genetics, College of Computer, Mathematics and Natural Sciences, University of Maryland, College Park, MD, USA.
Matthias
M
Corrotte
Maria Cecilia
MC
Fernandes
Christina
C
Tam
Norma W
NW
Andrews
D000818:Animals; D001426:Bacterial Proteins / Q000378:metabolism; D001427:Bacterial Toxins / Q000378:metabolism*; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism*; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D004705:Endocytosis; D008954:Models, Biological; D057146:Multivesicular Bodies / Q000378:metabolism*; D013301:Streptolysins / Q000378:metabolism
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D013301:Streptolysins; C025964:streptolysin O