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PMID: 11937624 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

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

Nucleic acids research ·Vol. 30 ·No. 8 ·2002-04-15 ·Pages 1713-24

Braastad CD, Leguia M, Hendrickson EA

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 approximately 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.

MeSH Terms
5' Flanking Region Base Sequence Carrier Proteins/biosynthesis,genetics CpG Islands DNA Helicases DNA Methylation Deoxyribonuclease I/chemistry Fatty Acid Desaturases/biosynthesis,genetics Humans Ku Autoantigen Molecular Sequence Data NADH, NADPH Oxidoreductases Oxidoreductases Acting on CH-CH Group Donors RNA, Messenger/biosynthesis Radiation, Ionizing Response Elements Transcription Initiation Site Transcriptional Activation Tumor Cells, Cultured Tumor Suppressor Protein p53/metabolism
Chemicals
Carrier Proteins RNA, Messenger Tumor Suppressor Protein p53 Fatty Acid Desaturases Oxidoreductases Acting on CH-CH Group Donors trans-2-enoyl-CoA reductase (NADPH) NADH, NADPH Oxidoreductases Deoxyribonuclease I DNA Helicases XRCC5 protein, human Ku Autoantigen
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Braastad Corey D
Department of Molecular Biology, Cellular Biology and Biochemistry, Brown University, Providence, RI 02912, USA.
Leguia Mariana
Hendrickson Eric A
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/30.8.1713
Published
2002-04-15
Pages
1713-24
Language
English
Region
England
NLM ID
0411011
PMCID
PMC113227
Subset
IM
Grants
NIAID NIH HHS · AI35763 · United States
External Links
PubMed source DOI: 10.1093/nar/30.8.1713 PMC Full Text View journal details
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