Regulation of sympathetic neuron and neuroblastoma cell death by XIAP and its association with proteasomes in neural cells.
XIAP (X chromosome-linked inhibitor of apoptosis protein) has been shown to inhibit cell death in a variety of cells. XIAP binds to active caspases, but XIAP also has a carboxy-terminal RING domain that can regulate cell death via protein degradation. Here we have studied the function of full-length and RING-deleted XIAP in mouse sympathetic neurons microinjected with expression plasmids and in neuroblastoma cells stably overexpressing these proteins. Both full-length and RING-deleted XIAP-protected sympathetic neurons against death induced by nerve growth factor (NGF) withdrawal to about the same extent. However, the two proteins were differentially localized in transfected neurons, with RING-deleted XIAP present in the cytoplasm and full-length XIAP found mostly in cytoplasmic protein aggregates, as revealed by transmission electron microscopy. The occurrence of these aggregates was blocked by lactacystin, a proteasome inhibitor. In neuroblastoma cells, RING-deleted XIAP protected against death induced by staurosporine, thapsigargin, or serum withdrawal, whereas full-length XIAP was without effect. Full-length, but not RING-deleted, XIAP was degraded and ubiquitinated in the neuroblastoma cells. The results show that the presence of the RING domain differentially affected the neuroprotective ability of XIAP in sensory neurons and neuroblastoma cells. The RING domain was essentially required for the proteasomal association of XIAP and for its ubiquitination.
Yu
Li-Ying
LY
Program of Molecular Neurobiology, Institute of Biotechnology, University of Helsinki, Finland.
Korhonen
Laura
L
Martinez
Rodrigo
R
Jokitalo
Eija
E
Chen
Yuming
Y
Arumäe
Urmas
U
Lindholm
Dan
D
D000818:Animals; D019556:COS Cells; D016923:Cell Death / Q000502:physiology*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D017353:Gene Deletion; D006801:Humans; D051379:Mice; D008815:Mice, Inbred Strains; D009097:Multienzyme Complexes / Q000378:metabolism*; D016296:Mutagenesis; D009447:Neuroblastoma; D009474:Neurons / Q000166:cytology* / Q000201:enzymology*; D046988:Proteasome Endopeptidase Complex; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017783:Superior Cervical Ganglion / Q000166:cytology; D014407:Tumor Cells, Cultured; D025801:Ubiquitin / Q000378:metabolism; D051636:X-Linked Inhibitor of Apoptosis Protein
D009097:Multienzyme Complexes; D011506:Proteins; D025801:Ubiquitin; D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex