signature=d6c77c3cb1618254bc41c02c41c3eeac,The Retinoic Acid Receptor ␣ (RAR␣) Chimeric Proteins PML...

摘要：

Retinoic acid receptor (RAR) gene rearrangementby reciprocal chromosome translocation is themolecular signature of acute promyelocytic leukemia(APL). Disruption of RAR function appears to be thelikely cause of aberrant myelopoiesis observed in APL, because PML-RAR expression has been shown toderegulate the transcription of genes that controlmyelopoiesis. To target RAR chimeric proteins, weengineered epitope-tagged versions of PML-RAR,PLZF-RAR, NPM-RAR, and NuMA-RAR (X-RARV5) and generated a panel of stable COS cell linesexpressing X-RARV5. Protein fractionation andWestern analysis of these COS lines reveal thatX-RAR proteins localize to both the cytoplasm andnucleus. NPM-RAR is predominantly nuclear whereas NuMA-RAR is predominantly cytoplasmic. Confocalimmunofluorescent microscopy reveals that PML-RAR and PLZF-RAR share a primarily diffuse nuclear pattern that excludes the nucleolus. NPM-RAR is also diffuse in the nucleus but, in contrast to PML-RARand PLZF-RAR, is strongly associated with thenucleolus. Strikingly, NuMA-RAR predominantlylocalizes throughout the cytoplasm in a microspeckled pattern. We further demonstrate that NPM and NuMA interact with NPM-RAR and NuMA-RAR,respectively. The distinct intracellular localizationpatterns and the shared ability of X-RAR to interactwith their respective RAR partner proteins (X) further support the hypothesis that deregulation of thesepartners may play a role in APL pathogenesis.

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