UBE1L represses PML/RAR{alpha} by targeting the PML domain for ISG15ylation

Mol Cancer Ther. 2008 Apr;7(4):905-14. doi: 10.1158/1535-7163.MCT-07-0515.

Abstract

Acute promyelocytic leukemia (APL) is characterized by expression of promyelocytic leukemia (PML)/retinoic acid (RA) receptor alpha (RARalpha) protein and all-trans-RA-mediated clinical remissions. RA treatment can confer PML/RARalpha degradation, overcoming dominant-negative effects of this oncogenic protein. The present study uncovered independent retinoid degradation mechanisms, targeting different domains of PML/RARalpha. RA treatment is known to repress PML/RARalpha and augment ubiquitin-activating enzyme-E1-like (UBE1L) protein expression in NB4-S1 APL cells. We previously reported RA-induced UBE1L and the IFN-stimulated gene, 15-kDa protein ISG15ylation in APL cells. Whether the ubiquitin-like protein ISG15 directly conjugates with PML/RARalpha was not explored previously and is examined in this study. Transient transfection experiments with different PML/RARalpha domains revealed that RA treatment preferentially down-regulated the RARalpha domain, whereas UBE1L targeted the PML domain for repression. As expected, ubiquitin-specific protease 18 (UBP43/USP18), the ISG15 deconjugase, opposed UBE1L but not RA-dependent PML/RARalpha degradation. In contrast, the proteasomal inhibitor, N-acetyl-leucinyl-leucinyl-norleucinal, inhibited both UBE1L- and RA-mediated PML/RARalpha degradation. Notably, UBE1L induced ISG15ylation of the PML domain of PML/RARalpha, causing its repression. These findings confirmed that RA triggers PML/RARalpha degradation through different domains and distinct mechanisms. Taken together, these findings advance prior work by establishing two pathways converge on the same oncogenic protein to cause its degradation and thereby promote antineoplastic effects. The molecular pharmacologic implications of these findings are discussed.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Bronchi / cytology
  • Bronchi / metabolism
  • COS Cells
  • Cells, Cultured
  • Chlorocebus aethiops
  • Cytokines / metabolism*
  • Endopeptidases / metabolism
  • Gene Expression Regulation, Leukemic*
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Leukemia, Promyelocytic, Acute / genetics
  • Leukemia, Promyelocytic, Acute / metabolism*
  • Leukemia, Promyelocytic, Acute / pathology
  • Leupeptins / pharmacology
  • Oncogene Proteins, Fusion / antagonists & inhibitors
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism*
  • Protein Processing, Post-Translational
  • Protein Structure, Tertiary
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Signal Transduction
  • Transfection
  • Tretinoin / pharmacology
  • Ubiquitin Thiolesterase
  • Ubiquitin-Activating Enzymes / pharmacology*
  • Ubiquitins / metabolism*

Substances

  • Antineoplastic Agents
  • Cytokines
  • Leupeptins
  • Oncogene Proteins, Fusion
  • RNA, Messenger
  • Ubiquitins
  • promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein
  • acetylleucyl-leucyl-norleucinal
  • Tretinoin
  • ISG15 protein, human
  • Endopeptidases
  • USP18 protein, human
  • Ubiquitin Thiolesterase
  • Ubiquitin-Activating Enzymes