Ku86 autoantigen related protein-1 transcription initiates from a CpG island and is induced by p53 through a nearby p53 response element

Corey D. Braastad, Mariana Leguia, Eric A. Hendrickson

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

The human Ku86 gene and an isoform, KARP-1 (Ku86 autoantigen related protein-1), encode overlapping, but differentially regulated, transcripts. Ku86 is constitutively transcribed at high levels and, although it plays a seminal role in DNA double-strand break repair, its expression is not induced by DNA damage. KARP-1, in contrast, is expressed constitutively only at low levels and its expression is induced by DNA damage in a p53-dependent fashion. The regulatory elements promoting KARP-1 gene expression and p53 responsiveness, however, were unknown. Here, we report that a strong DNase I hypersensitive site (DHS) resides ∼25 kb upstream from the Ku86 promoter. This DHS is encompassed by a hypomethylated CpG island. Reporter assays demonstrated that this region corresponded to a promoter(s), which promoted transcription of peroxisomal trans-2-enoyl CoA reductase in the centromeric direction and KARP-1 in the telomeric direction. KARP-1 primer extension products were mapped to this CpG island in the correct transcriptional orientation confirming that KARP-1 transcription initiates from this site. Moreover, a p53 response element within the first intron of the KARP-1 transcriptional unit was identified using chromatin immunoprecipitation and antibodies specific to activated forms of p53. These data expand our understanding of this important DNA repair locus.

Original languageEnglish (US)
Pages (from-to)1713-1724
Number of pages12
JournalNucleic acids research
Volume30
Issue number8
DOIs
StatePublished - Apr 15 2002

Bibliographical note

Funding Information:
We are deeply indebted to Drs Ken Zaret, Anja-Katrin Bielinsky and Ms Jaqueline Brooks for their advice and help with the DNase I hypersensitivity, chromatin immunoprecipitation assays and 5′-RACE protocols, respectively. We thank Dr Bielinsky for her comments and criticisms of the manuscript. This work was supported in part by a grant from the NIH (AI35763).

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