TY - JOUR
T1 - Transcriptional regulation of the p67phox gene
T2 - Role of AP-1 in concert with myeloid-specific transcription factors
AU - Li, Sen Lin
AU - Valente, Anthony J
AU - Wang, Long
AU - Gamez, Maria J.
AU - Clark, Robert A.
PY - 2001/10/19
Y1 - 2001/10/19
N2 - 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.
AB - 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.
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U2 - 10.1074/jbc.M106111200
DO - 10.1074/jbc.M106111200
M3 - Article
C2 - 11483614
AN - SCOPUS:0035914325
SN - 0021-9258
VL - 276
SP - 39368
EP - 39378
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 42
ER -