Formation of PML/RAR alpha high molecular weight nuclear complexes through the PML coiled-coil region is essential for the PML/RAR alpha-mediated retinoic acid response

Oncogene. 1999 Nov 4;18(46):6313-21. doi: 10.1038/sj.onc.1203029.

Abstract

Retinoic Acid (RA) treatment induces disease remission of Acute Promyelocytic Leukaemia (APL) patients by triggering terminal differentiation of neoplastic cells. RA-sensitivity in APL is mediated by its oncogenic protein, which results from the recombination of the PML and the RA receptor alpha (RAR alpha) genes (PML/RAR alpha fusion protein). Ectopic expression of PML/RAR alpha into haemopoietic cell lines results in increased response to RA-induced differentiation. By structure-function analysis of PML/RAR alpha-mediated RA-differentiation, we demonstrated that fusion of PML and RAR alpha sequences and integrity of the PML dimerization domain and of the RAR alpha DNA binding region are required for the effect of PML/RAR alpha on RA-differentiation. Indeed, direct fusion of the PML dimerization domain to the N- or C-terminal extremities of RAR alpha retained full biological activity. All the biologically active PML/RAR alpha mutants formed high molecular weight complexes in vivo. Functional analysis of mutations within the PML dimerization domain revealed that the capacity to form PML/RAR alpha homodimers, but not PML/RAR alpha-PML heterodimers, correlated with the RA-response. These results suggest that targeting of RAR alpha sequences by the PML dimerization domain and formation of nuclear PML/RAR alpha homodimeric complexes are crucial for the ability of PML/RAR alpha to mediate RA-response.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antineoplastic Agents / pharmacology*
  • Binding Sites
  • Cell Differentiation / drug effects
  • DNA, Neoplasm / genetics
  • DNA, Neoplasm / metabolism
  • Dimerization
  • Gene Expression Regulation, Neoplastic / drug effects
  • HeLa Cells / drug effects
  • Hematopoietic Stem Cells / cytology
  • Hematopoietic Stem Cells / drug effects
  • Hematopoietic Stem Cells / metabolism
  • Humans
  • Leukemia, Promyelocytic, Acute / drug therapy
  • Leukemia, Promyelocytic, Acute / genetics*
  • Leukemia, Promyelocytic, Acute / metabolism
  • Molecular Weight
  • Neoplasm Proteins / chemistry*
  • Neoplasm Proteins / physiology
  • Oncogene Proteins, Fusion / chemistry*
  • Oncogene Proteins, Fusion / physiology
  • Protein Structure, Tertiary
  • Recombinant Fusion Proteins / metabolism
  • Structure-Activity Relationship
  • Tretinoin / pharmacology*
  • U937 Cells / drug effects
  • U937 Cells / metabolism
  • Zinc Fingers

Substances

  • Antineoplastic Agents
  • DNA, Neoplasm
  • Neoplasm Proteins
  • Oncogene Proteins, Fusion
  • Recombinant Fusion Proteins
  • promyelocytic leukemia-retinoic acid receptor alpha fusion oncoprotein
  • Tretinoin