Promoter function of the human ap-endonuclease

T. Izumi, W. D. Henner, Sankar Mitra

Research output: Contribution to journalArticle

Abstract

The cDNA of the major apurinic/apyrimidinic endonuclease (APE), named APE-1, has been cloned, not only on the basis of its nuclease activity in base excision repair pathway but also because of its role as an activator of AP-1 transcription factor and as a represser of parathyroid hormone (PTH) gene. APE-1 was shown to bind two negative Ca2+-dependent response elements (nCaRE) in the PTH promoter. In a systematic effort to characterize regulatory elements in the APE-1 promoter, we constructed a series of expression plasmids in which the luciferase coding sequence is fused to overlapping segments of the APE-1 promoter. Transient expression studies of luciferase in electroporated human cells showed -10-fold lower expression of the reporter from plasmids containing a 3.9 kb promoter than that from a plasmid in which a 2.0 kb upstream fragment was deleted. This result suggested the presence of a negative regulatory sequence in the upstream region. A more detailed deletion analysis of the reporter plasmid narrowed the activity to a 800 bp region that is needed for repression of luciferase. Sequencing of this DNA region reveals the presence of an Alu sequence and two nCaRE sequences. Because APE-1 binds to nCARE, the possibility of its feedback autoregulation is being investigated.

Original languageEnglish (US)
JournalFASEB Journal
Volume10
Issue number6
StatePublished - Dec 1 1996

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Genetics

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