Transcriptional regulation of the p67phox gene: Role of AP-1 in concert with myeloid-specific transcription factors

Sen Lin Li, Anthony J. Valente, Long Wang, Maria J. Gamez, Robert A. Clark

Research output: Contribution to journalArticle

23 Scopus citations

Abstract

We have investigated the myeloid-specific transcriptional regulation of p67phox an essential component of phagocyte respiratory burst NADPH oxidase. Analysis was carried out on the p67phox 5′-flanking region from -3669 to -4 (relative to ATG), including the first exon and intron and part of the second exon. The construct extending from -985 to -4 produced the highest luciferase activity in myeloid HL-60 cells but was not active in HeLa or Jurkat cells, indicating myeloid-specific expression. Four active elements were identified: Sp1/Sp3 at -694, PU.1 at -289, AP-1 at -210, and PU.1/HAF1 at -182, the latter three being in the first intron. These cis elements bound their cognate transacting factors both in vitro and in vivo. Mutation of the Sp1, PU.1, or PU.1/ HAF1 site each decreased promoter activity by 35-50%. Mutations in all three sites reduced promoter activity by 90%. However, mutation of the AP-1 site alone nearly abolished promoter activity. The AP-1 site bound Jun and Fos proteins from HL-60 cell nuclear extract. Coexpression with Jun B in AP-1-deficient cells increased promoter activity by 3-fold. These data show that full p67phox promoter activity requires cooperation between myeloid-specific and nonmyeloid transcription factors, with AP-1 being the most critical for function.

Original languageEnglish (US)
Pages (from-to)39368-39378
Number of pages11
JournalJournal of Biological Chemistry
Volume276
Issue number42
DOIs
Publication statusPublished - Oct 19 2001

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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