Functional analysis of the transcription factor ER71 and its activation of the matrix metalloproteinase-1 promoter

Nucleic Acids Res. 2002 Jul 1;30(13):2972-9. doi: 10.1093/nar/gkf390.

Abstract

The ETS transcription factor family is characterized by a conserved ETS DNA-binding domain and its members have been implicated in a plethora of biological processes, including development, cell transformation and metastasis. ER71 is a testis-specific ETS protein that is not homologous to any other protein outside its ETS domain, suggesting that it fulfills a unique physiological role. Here, we report that ER71 is a constitutively nuclear protein whose intracellular localization is dependent on a portion of the ETS domain, namely ER71 amino acids 276-315. Furthermore, the DNA binding activity is intramolecularly regulated, as the N-terminus of ER71 has a negative effect on DNA binding while the C-terminus dramatically enhances this activity. We also demonstrate that ER71 possesses an extremely potent N-terminal transactivation domain comprised of amino acids 1-157. Finally, we show that ER71 is capable of directly activating both an E74 site-driven and the matrix metalloproteinase-1 promoter. Altogether, these data represent the first functional characterization of ER71, which may perform important functions in the developing and adult testis as well as in testicular germ cell tumorigenesis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites / genetics
  • Binding, Competitive
  • Cell Line
  • Cell Nucleolus / metabolism
  • Cell Nucleus / metabolism
  • Electrophoretic Mobility Shift Assay
  • Gene Expression Regulation
  • Green Fluorescent Proteins
  • Humans
  • Luciferases / genetics
  • Luciferases / metabolism
  • Luminescent Proteins / genetics
  • Luminescent Proteins / metabolism
  • Matrix Metalloproteinase 1 / genetics*
  • Microscopy, Fluorescence
  • Oligonucleotides / genetics
  • Oligonucleotides / metabolism
  • Promoter Regions, Genetic / genetics*
  • Protein Binding
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transcription Factors / genetics
  • Transcription Factors / physiology*
  • Transcription, Genetic
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Luminescent Proteins
  • Oligonucleotides
  • Recombinant Fusion Proteins
  • Transcription Factors
  • Green Fluorescent Proteins
  • Luciferases
  • Matrix Metalloproteinase 1