Identification of a DNA-binding site and transcriptional target for the EWS-WT1(+KTS) oncoprotein

Paul Andrew Reynolds, GA Smolen, RE Palmer, D Sgroi, V Yajnik, WL Gerald, DA Haber

Research output: Contribution to journalArticlepeer-review

41 Citations (Scopus)

Abstract

Desmoplastic small round cell tumor (DSRCT) is defined by a chimeric transcription factor, resulting from fusion of the N-terminal domain of the Ewing's sarcoma gene EWS to the three C-terminal zinc fingers of the Wilms' tumor suppressor WT1. Although DNA-binding sites have been defined for the uninterrupted WT1 zinc finger domains, the most prevalent isoforms of both WT1 and EWS-WT1 have an insertion of three amino acids [lysine, threonine, and serine (KTS)], which abrogates binding to known consensus sequences and transactivation of known target genes. Here, we used cDNA subtractive hybridization to identify an endogenous gene, LRRC15, which is specifically up-regulated after inducible expression of EWS-WT1(+KTS) in cancer cell lines, and is expressed within primary DSRCT cells. The chimeric protein binds in vitro and in vivo to a specific element upstream of LRRC15, leading to dramatic transcriptional activation. Mutagenesis studies define the optimal binding site of the (+KTS) isoform of EWS-WT1 as 5'-GGAGG(A/G)-3'. LRRC15 encodes a leucine-rich transmembrane protein, present at the leading edge of migrating cells, the expression of which in normal tissues is restricted to the invasive cytotrophoblast layer of the placenta; small interfering (siRNA)-mediated suppression of LRRC15 expression in breast cancer cells leads to abrogation of invasiveness in vitro. Together, these observations define the consequence of (KTS) insertion within WT1-derived zinc fingers, and identify a novel EWS-WT1 transcriptional target implicated in tumor invasiveness.

Original languageEnglish
Pages (from-to)2094-2107
Number of pages14
JournalGenes & Development
Volume17
Issue number17
DOIs
Publication statusPublished - 1 Sept 2003

Keywords

  • DSRCT
  • EWS-WT1
  • alternative splicing
  • promoter
  • zinc finger
  • invasion
  • ROUND-CELL TUMOR
  • ADJACENT ZINC FINGERS
  • LEUCINE-RICH REPEAT
  • WILMS-TUMOR
  • TRANSLOCATION PRODUCT
  • FRASIER-SYNDROME
  • PROSTATE-CANCER
  • REACTIVE STROMA
  • FUSION PROTEIN
  • HUMAN GENOME

Fingerprint

Dive into the research topics of 'Identification of a DNA-binding site and transcriptional target for the EWS-WT1(+KTS) oncoprotein'. Together they form a unique fingerprint.

Cite this