The SQSTM1-NUP214 fusion protein interacts with Crm1, activates Hoxa and Meis1 genes, and drives leukemogenesis in mice

PLoS One. 2020 Apr 28;15(4):e0232036. doi: 10.1371/journal.pone.0232036. eCollection 2020.

Abstract

The NUP98 and NUP214 nucleoporins (NUPs) are recurrently fused to heterologous proteins in leukemia. The resulting chimeric oncoproteins retain the phenylalanine-glycine (FG) repeat motifs of the NUP moiety that mediate interaction with the nuclear export receptor Crm1. NUP fusion leukemias are characterized by HOXA gene upregulation; however, their molecular pathogenesis remains poorly understood. To investigate the role of Crm1 in mediating the leukemogenic properties of NUP chimeric proteins, we took advantage of the Sequestosome-1 (SQSTM1)-NUP214 fusion. SQSTM1-NUP214 retains only a short C-terminal portion of NUP214 which contains FG motifs that mediate interaction with Crm1. We introduced point mutations targeting these FG motifs and found that the ability of the resulting SQSTM1-NUP214FGmut protein to interact with Crm1 was reduced by more than 50% compared with SQSTM1-NUP214. Mutation of FG motifs affected transforming potential: while SQSTM1-NUP214 impaired myeloid maturation and conferred robust colony formation to transduced hematopoietic progenitors in a serial replating assay, the effect of SQSTM1-NUP214FGmut was considerably diminished. Moreover, SQSTM1-NUP214 caused myeloid leukemia in all transplanted mice, whereas none of the SQSTM1-NUP214FGmut reconstituted mice developed leukemia. These oncogenic effects coincided with the ability of SQSTM1-NUP214 and SQSTM1-NUP214FGmut to upregulate the expression of Hoxa and Meis1 genes in hematopoietic progenitors. Indeed, chromatin immunoprecipitation assays demonstrated that impaired SQSTM1-NUP214 interaction with Crm1 correlated with impaired binding of the fusion protein to Hoxa and Meis1 genes. These findings highlight the importance of Crm1 in mediating the leukemogenic properties of SQSTM1-NUP214, and suggest a conserved role of Crm1 in recruiting oncoproteins to their effector genes.

Publication types

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

MeSH terms

  • Amino Acid Motifs
  • Animals
  • Cell Line
  • Exportin 1 Protein
  • Gene Expression Regulation, Neoplastic
  • Homeodomain Proteins / genetics*
  • Karyopherins / metabolism*
  • Leukemia / genetics
  • Leukemia / metabolism*
  • Leukemia / pathology
  • Mice
  • Mutagenesis, Site-Directed
  • Myeloid Ecotropic Viral Integration Site 1 Protein / genetics*
  • Neoplasm Transplantation
  • Nuclear Pore Complex Proteins / chemistry
  • Nuclear Pore Complex Proteins / genetics*
  • Oncogene Proteins, Fusion / chemistry
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism*
  • Point Mutation
  • Receptors, Cytoplasmic and Nuclear / metabolism*
  • Sequestosome-1 Protein / genetics*
  • Up-Regulation

Substances

  • Homeodomain Proteins
  • Karyopherins
  • Meis1 protein, mouse
  • Myeloid Ecotropic Viral Integration Site 1 Protein
  • Nuclear Pore Complex Proteins
  • Nup214 protein, mouse
  • Oncogene Proteins, Fusion
  • Receptors, Cytoplasmic and Nuclear
  • Sequestosome-1 Protein
  • Sqstm1 protein, mouse
  • HoxA protein