Mass spectroscopy identifies the splicing-associated proteins, PSF, hnRNP H3, hnRNP A2/B1, and TLS/FUS as interacting partners of the ZNF198 protein associated with rearrangement in myeloproliferative disease

Exp Cell Res. 2005 Sep 10;309(1):78-85. doi: 10.1016/j.yexcr.2005.05.019.

Abstract

ZNF198 is fused with FGFR1 in an atypical myeloproliferative disease that results in constitutive activation of the kinase domain and mislocalization to the cytoplasm. We have used immunoprecipitation of a GFP-tagged ZNF198 combined with MALDI-TOF mass spectroscopy to identify interacting proteins. P splicing factor (PSF) was identified as one of the proteins and this interaction was confirmed by Western blotting. Other proteins identified were the spliceosomal components hnRNP A2/B1, hnRNP H3, and TLS/FUS. PSF is also known to interact with PTB, another member of the hnRNP family of proteins, and we further demonstrated that PTB interacts with ZNF198. The interaction between TLS/FUS and ZNF198 was confirmed using Western blot analysis. In 293 cells expressing the ZNF198/FGFR1 fusion protein, neither PSF nor PTB binds to the fusion protein, possibly because of their differential localization in the cell.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Carrier Proteins* / chemistry
  • Carrier Proteins* / genetics
  • Carrier Proteins* / metabolism
  • Cell Line
  • DNA-Binding Proteins* / chemistry
  • DNA-Binding Proteins* / genetics
  • DNA-Binding Proteins* / metabolism
  • Gene Rearrangement
  • Humans
  • Mass Spectrometry
  • Molecular Sequence Data
  • Myeloproliferative Disorders / genetics
  • Myeloproliferative Disorders / metabolism*
  • Oncogene Proteins, Fusion / genetics
  • PTB-Associated Splicing Factor
  • Protein Binding
  • Proto-Oncogene Proteins / genetics
  • RNA Splicing / genetics
  • RNA-Binding Protein FUS / chemistry
  • RNA-Binding Protein FUS / genetics
  • RNA-Binding Protein FUS / metabolism
  • RNA-Binding Proteins* / chemistry
  • RNA-Binding Proteins* / genetics
  • RNA-Binding Proteins* / metabolism
  • Ribonucleoproteins, Small Nuclear / chemistry
  • Ribonucleoproteins, Small Nuclear / genetics
  • Ribonucleoproteins, Small Nuclear / metabolism
  • Transcription Factors

Substances

  • CEP43 protein, human
  • Carrier Proteins
  • DNA-Binding Proteins
  • Oncogene Proteins, Fusion
  • PTB-Associated Splicing Factor
  • Proto-Oncogene Proteins
  • RNA-Binding Protein FUS
  • RNA-Binding Proteins
  • Ribonucleoproteins, Small Nuclear
  • Transcription Factors
  • ZMYM2 protein, human