Transcriptional regulation of the p67phox gene

Role of AP-1 in concert with myeloid-specific transcription factors

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

Research output: Contribution to journalArticle

23 Citations (Scopus)

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
StatePublished - Oct 19 2001

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Transcription Factor AP-1
Transcription Factors
Genes
HL-60 Cells
Introns
Mutation
Exons
Jurkat Cells
NADPH Oxidase
5' Flanking Region
Myeloid Cells
Phagocytes
Cell Extracts
Luciferases
HeLa Cells
neutrophil cytosol factor 67K
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Transcriptional regulation of the p67phox gene : Role of AP-1 in concert with myeloid-specific transcription factors. / Li, Senlin; Valente, Anthony J.; Wang, Long; Gamez, Maria J.; Clark, Robert A.

In: Journal of Biological Chemistry, Vol. 276, No. 42, 19.10.2001, p. 39368-39378.

Research output: Contribution to journalArticle

Li, Senlin ; Valente, Anthony J. ; Wang, Long ; Gamez, Maria J. ; Clark, Robert A. / Transcriptional regulation of the p67phox gene : Role of AP-1 in concert with myeloid-specific transcription factors. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 42. pp. 39368-39378.
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