title: The acute promyelocytic leukaemia specific PML and PLZF proteins localize to adjacent and functionally distinct nuclear bodies.
abstract: Acute promyelocytic leukaemia is characterized by translocations that involve the retinoic acid receptor alpha (RAR alpha) locus on chromosome 17 and the PML locus on 15 or the PLZF locus on 11. The resulting abnormal translocation products encode for PML/RAR alpha or PLZF/RAR alpha fusion proteins. There is increasing experimental evidence that the APL-specific fusion proteins have similar biologic activities on differentiation and survival and that both components of the fusion proteins (PML or PLZF and RAR alpha) are indispensable for these biological activities. The physiologic function of PML or PLZF or whether PML and PLZF contribute common structural or functional features to the corresponding fusion proteins is not known. We report here immunofluorescence studies on the cellular localization of PLZF and PLZF/RAR alpha and compare it with the localization of PML and PML/RAR alpha. PLZF localizes to nuclear domains of 0.3-0.5 microns, approximately 14 per cell in the KG1 myeloid cell line. These PLZF-bodies are morphologically similar to the domains reported for PML (PML-NBs). There is tight spatial relationship between about 30% of PLZ-NBs and PML-NBs: they partially overlap. However, PML and PLZF do not form soluble complexes in vivo. PLZF- and PML-NBs are functionally distinct. Adenovirus E4-ORF3 protein expression alters the structure of the PML-NBs and interferon increases the number of PML-NBs and neither has any effect on PLZF NBs. The localization of PLZF/RAR alpha is different to that of PLZF and RAR alpha. The nuclear distribution pattern of PLZF/RAR alpha is one of hundreds of small dots (microspeckles) less than 0.1 micron. Expression of PLZF/RAR alpha did not provoke disruption of the PML-NBs. Co-expression of PML/RAR alpha and PLZF/RAR alpha in U937 cells revealed apparent colocalization. Overall the results suggest that the PML- and PLZF-NBs are distinct functional nuclear domains, but that they may share common regulatory pathways and/or targeting sequences, as revealed by the common localization of their corresponding fusion proteins.
authors.affiliation: European Institute of Oncology, Department of Experimental Oncology, Milan, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Ruthardt
authors.forename: A
authors.initials: A
authors.lastname: Orleth
authors.forename: L
authors.initials: L
authors.lastname: Tomassoni
authors.forename: E
authors.initials: E
authors.lastname: Puccetti
authors.forename: D
authors.initials: D
authors.lastname: Riganelli
authors.forename: M
authors.initials: M
authors.lastname: Alcalay
authors.forename: R
authors.initials: R
authors.lastname: Mannucci
authors.forename: I
authors.initials: I
authors.lastname: Nicoletti
authors.forename: F
authors.initials: F
authors.lastname: Grignani
authors.forename: M
authors.initials: M
authors.lastname: Fagioli
authors.forename: P G
authors.initials: PG
authors.lastname: Pelicci
mesh_terms: D002467:Cell Nucleus / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D051741:Kruppel-Like Transcription Factors; D009363:Neoplasm Proteins / Q000032:analysis*; D009687:Nuclear Proteins / Q000032:analysis*; D000071156:Promyelocytic Leukemia Protein; D000075722:Promyelocytic Leukemia Zinc Finger Protein; D018168:Receptors, Retinoic Acid / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000032:analysis; D000072482:Retinoic Acid Receptor alpha; D014157:Transcription Factors / Q000032:analysis*; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins
chemical_list: D004268:DNA-Binding Proteins; D051741:Kruppel-Like Transcription Factors; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D000071156:Promyelocytic Leukemia Protein; D000075722:Promyelocytic Leukemia Zinc Finger Protein; C000607071:RARA protein, human; D018168:Receptors, Retinoic Acid; D011993:Recombinant Fusion Proteins; D000072482:Retinoic Acid Receptor alpha; D014157:Transcription Factors; D025521:Tumor Suppressor Proteins; C070576:PML protein, human; C092037:ZBTB16 protein, human