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Two evolutionarily conserved sequence elements for Peg3/Usp29 transcription.

Jeong Do Kim1, Sungryul Yu, Jung Ha Choo

  • 1Department of Biological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA. jkim@lsu.edu

BMC Molecular Biology
|December 11, 2008
PubMed
Summary
This summary is machine-generated.

Conserved Sequence Elements CSE1 and CSE2 regulate imprinted gene expression. CSE2 activates Peg3 transcription, while CSE1 represses both Peg3 and Usp29 transcription.

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Area of Science:

  • Genetics
  • Epigenetics
  • Molecular Biology

Background:

  • The Peg3 imprinted domain contains two imprinted genes, Peg3 and Usp29, regulated by a bidirectional promoter within a CpG island.
  • This region contains two Conserved Sequence Elements, CSE1 and CSE2, which are YY1 binding sites and are hypothesized to function as an Imprinting Control Region (ICR).

Purpose of the Study:

  • To investigate the functional roles of CSE1 and CSE2 in the transcriptional regulation of Peg3 and Usp29.
  • To determine the specific roles of YY1 binding sites within CSE2 and the repressive regions within CSE1.

Main Methods:

  • Cell line-based promoter assays were employed to assess the transcriptional activity.
  • Site-directed mutagenesis was used to mutate CSE1 and CSE2 binding sites.
  • Serial mutational analyses were performed to pinpoint critical regions within CSE1.

Main Results:

  • Mutation of CSE2 (YY1 binding sites) reduced Peg3 promoter activity in an orientation-dependent manner, indicating an activator role for CSE2 in Peg3 transcription.
  • CSE2 mutation did not affect Usp29 promoter activity.
  • Mutation of CSE1 increased promoter activity for both Peg3 and Usp29 in an orientation-dependent manner, suggesting a repressor role.
  • Further analysis identified two distinct 6-bp regions within CSE1 responsible for repressing Peg3 and Usp29 transcription.

Conclusions:

  • CSE2 functions as a transcriptional activator for the Peg3 gene.
  • CSE1 acts as a transcriptional repressor for both Peg3 and Usp29 genes.
  • These findings elucidate the distinct roles of CSE1 and CSE2 in regulating the Peg3 imprinted domain.