The KRAB-containing zinc-finger transcriptional regulator ZBRK1 activates SCA2 gene transcription through direct interaction with its gene product, ataxin-2

Hum Mol Genet. 2011 Jan 1;20(1):104-14. doi: 10.1093/hmg/ddq436. Epub 2010 Oct 6.

Abstract

Gene transcription is controlled by transcriptional regulators acting with specific co-regulators to allow gene activation and repression. Here, we report the identification of the KRAB-containing zinc-finger transcriptional regulator, ZBRK1, as an interaction partner of the SCA2 gene product ataxin-2. Furthermore, we discovered that an elevated ZBRK1 level resulted in increased ataxin-2 levels, whereas interference on transcriptional and protein levels of ZBRK1 yielded reduced ataxin-2 levels, suggesting that a complex comprising ZBRK1 and ataxin-2 regulates SCA2 gene transcription. A bioinformatic analysis utilizing the known ZBRK1 consensus DNA-binding motif revealed ZBRK1-binding sites in the SCA2 promoter. These predicted sites were experimentally validated by chromatin-immunoprecipitation experiments along with luciferase-based promoter analyses corroborating that SCA2 gene transcription is controlled by a ZBRK1/ataxin-2 complex. Finally, we demonstrate that SCA2 gene transcription is significantly reduced in colon tumors possessing low ZBRK1 transcripts. Thus, our results provide first evidence that ataxin-2 acts as a co-regulator of ZBRK1 activating its own transcription, thereby representing the first identified ZBRK1 co-activator.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ataxins
  • Binding Sites
  • Chromatin / metabolism
  • Colonic Neoplasms / genetics
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Immunoprecipitation
  • Nerve Tissue Proteins / genetics*
  • Nerve Tissue Proteins / metabolism*
  • Plasmids
  • Promoter Regions, Genetic
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Spinocerebellar Ataxias / genetics*
  • Transcription, Genetic
  • Transcriptional Activation*

Substances

  • Ataxins
  • Chromatin
  • Nerve Tissue Proteins
  • Repressor Proteins
  • ZNF350 protein, human