Ubiquitin-activating enzyme, E1, is associated with maturation of autophagic vacuoles.
The ubiquitin-activating enzyme, E1, is required for initiating a multi-step pathway for the covalent linkage of ubiquitin to target proteins. A CHO cell line containing a mutant thermolabile E1, ts20, has been shown to be defective in stress-induced degradation of proteins at restrictive temperature (Gropper et al., 1991. J. Biol. Chem. 266:3602-3610). Parental E36 cells responded to restrictive temperature by stimulating lysosome-mediated protein degradation twofold. Such a response was not observed in ts20 cells. The absence of accelerated degradation in these cells at 39.5 degrees C was accompanied by an accumulation of autolysosomes. The fractional volume of these degradative autophagic vacuoles was at least sixfold greater than that observed for either E36 cells at 30.5 degrees or 39.5 degrees C, or ts20 cells at 30.5 degrees C. These vacuoles were acidic and contained both acid phosphatase and cathepsin L, but, unlike the autolysosomes observed in E36 cells, ubiquitin-conjugated proteins were conspicuously absent. Combined, our results suggest that in ts20 cells, which are unable to generate ubiquitin-protein conjugates due to heat inactivation of E1, the formation and maturation of autophagosomes into autolysosomes is normal, but the conversion of autolysosomes into residual bodies is disrupted.
Lenk
S E
SE
Department of Anatomy, University of Florida Health Science Center, Gainesville 32610.
Dunn
W A
WA
Trausch
J S
JS
Ciechanover
A
A
Schwartz
A L
AL
D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D001343:Autophagy; D016466:CHO Cells; D006224:Cricetinae; D008025:Ligases / Q000235:genetics / Q000378:metabolism*; D008247:Lysosomes / Q000502:physiology* / Q000648:ultrastructure; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D013696:Temperature; D044764:Ubiquitin-Activating Enzymes; D044767:Ubiquitin-Protein Ligases; D014452:Ubiquitins / Q000032:analysis / Q000378:metabolism; D014617:Vacuoles / Q000201:enzymology / Q000502:physiology* / Q000648:ultrastructure
D014452:Ubiquitins; D044767:Ubiquitin-Protein Ligases; D000135:Acid Phosphatase; D008025:Ligases; D044764:Ubiquitin-Activating Enzymes