Cloning and characterization of the novel chimeric gene p53/FXR2 in the acute megakaryoblastic leukemia cell line CMK11-5

Tohoku J Exp Med. 2006 Jul;209(3):169-80. doi: 10.1620/tjem.209.169.

Abstract

The loss of p53 function is a key event in tumorigenesis. Inactivation of p53 in primary tumors and cell lines is mediated by several molecular mechanisms, including deletions and rearrangements. However, generation of a p53 fusion gene has not yet been reported. Here we report a novel p53/an autosomal homolog of the fragile X mental retardation (FXR2) chimeric gene generated by an interstitial deletion. Western blot analyses have shown that the p53/FXR2 protein is indeed expressed in a Down syndrome-related acute megakaryoblastic leukemia cell line, CMK11-5 cells. To investigate the properties of the p53/FXR2 protein, we observed its subcellular localization. Flag-tagged expression vectors were transfected into COS-7 cells and the proteins were stained with an anti-Flag antibody. The p53/FXR2 protein was expressed at high levels in the cytoplasm, whereas wild-type p53 and FXR2 were localized primarily in the nucleus and in the periphery of the nucleus, respectively. Treatment with a topoisomerase II inhibitor, VP16, failed to induce expression of a p53 target gene, the cyclin-dependent kinase inhibitor p21(WAF-1/CIP1), in CMK11-5 cells, and transient transfection analysis showed that the p53/FXR2 protein failed to transactivate the p21(WAF-1/CIP1) promoter. These results suggest that the p53/FXR2 fusion protein lacks the ability of wild-type p53 to function as a transcription factor. The p53/FXR2 gene is the first reported p53 fusion gene.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • COS Cells
  • Cell Line, Tumor
  • Cell Transformation, Neoplastic
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Cyclin-Dependent Kinase Inhibitor p21 / metabolism
  • Etoposide / pharmacology
  • Gene Expression Profiling
  • Genes, p53 / genetics*
  • Humans
  • Leukemia, Megakaryoblastic, Acute / genetics*
  • Mice
  • Molecular Sequence Data
  • Mutant Chimeric Proteins / genetics*
  • NIH 3T3 Cells
  • RNA-Binding Proteins / genetics*
  • Topoisomerase II Inhibitors

Substances

  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • FXR2 protein, human
  • Mutant Chimeric Proteins
  • RNA-Binding Proteins
  • Topoisomerase II Inhibitors
  • Etoposide