MULTIVESICULAR ENDOSOMES CONTAINING INTERNALIZED EGF-EGF RECEPTOR COMPLEXES MATURE AND THEN FUSE DIRECTLY WITH LYSOSOMES

We have followed the transfer of EGF-EGF receptor (EGFR) complexes fro m endosomal vacuoles that contain transferrin receptors (TfR) to lysos ome vacuoles identified by their content of HRP loaded as a 15-min pul se 4 h previously. We show that the HRP-loaded lysosomes are lysosomal -associated membrane protein-1 (LAMP-1) positive, mannose-6-phosphate receptor (M6PR) negative, and contain active acid hydrolase. EGF-EGFR complexes are delivered to these lysosomes intact and are then rapidly degraded. Preactivating the HRP contained within the preloaded lysoso mes inhibits the delivery of EGFR and degradation of EGF, and results in the accumulation of EGFR-containing multivesicular bodies (MVB). Wi th time these accumulating MVB undergo a series of maturation changes that include the loss of TfR, the continued recruitment of EGFR, and t he accumulation of internal vesicles, but they remain LAMP-1 and M6PR negative, The mature MVB are often seen to make direct contact with ly sosomes containing preactivated HRP, but their perimeter membranes rem ain intact. Together our observations suggest that the transfer of EGF -EGFR complexes from the TfR-containing endosome compartment to the ly sosomes that degrade them employs a single vacuolar intermediate, the maturing MVB, and can be achieved by a single heterotypic fusion step.