Induction of AP-1 binding to intron 1 of SP-A1 and SP-A2 is implicated in the phorbol ester inhibition of human SP-A promoter activity

Russell R. Hoover, Jelena Pavlovic, Joanna Floros

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

A deletional analysis of the SP-A1 promoter in NCI-H441 cells was performed to identify potential cis-acting elements involved in phorbol ester-mediated repression of human SP-A transcription. The phorbol ester TPA reduced SP-A1 and SP-A2 promoter activity to approximately 35% to 45% compared to that of control cells. The inhibitory effect of TPA was significantly reduced upon removal of the region +64/+394 relative to the SP- A1 transcription start site. Using NCI-H441 nuclear proteins, electromobility shift assay analysis showed that the intron region +309/+329 of SP-A1 and the corresponding region of SP-A2 formed sequence-specific DNA/protein complexes that were induced by TPA exposure. The region +318/+324 of SP-A1 contains sequences similar to a consensus AP-1 binding site, TGACTGA (TCACTGA for SP- A2), which when mutated (TGAGAGT) prevented the formation of the TPA-induced DNA/protein complex. The TPA-induced complex was supershifted in the presence of antibody against the Fun family of proteins, but not the Fos family of proteins. These results suggest that the binding of AP-1 or an AP-1-like factor to the first intron of SP-A1 and SP-A2 may be involved in the phorbol ester inhibition of human SP-A gene expression.

Original languageEnglish (US)
Pages (from-to)303-317
Number of pages15
JournalExperimental Lung Research
Volume26
Issue number5
StatePublished - Jul 17 2000

Fingerprint

Transcription Factor AP-1
Phorbol Esters
varespladib methyl
Introns
Proteins
Transcription Initiation Site
DNA
Transcription
Nuclear Proteins
Gene expression
Assays
Binding Sites
Gene Expression
Antibodies

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Pulmonary and Respiratory Medicine
  • Clinical Biochemistry

Cite this

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title = "Induction of AP-1 binding to intron 1 of SP-A1 and SP-A2 is implicated in the phorbol ester inhibition of human SP-A promoter activity",
abstract = "A deletional analysis of the SP-A1 promoter in NCI-H441 cells was performed to identify potential cis-acting elements involved in phorbol ester-mediated repression of human SP-A transcription. The phorbol ester TPA reduced SP-A1 and SP-A2 promoter activity to approximately 35{\%} to 45{\%} compared to that of control cells. The inhibitory effect of TPA was significantly reduced upon removal of the region +64/+394 relative to the SP- A1 transcription start site. Using NCI-H441 nuclear proteins, electromobility shift assay analysis showed that the intron region +309/+329 of SP-A1 and the corresponding region of SP-A2 formed sequence-specific DNA/protein complexes that were induced by TPA exposure. The region +318/+324 of SP-A1 contains sequences similar to a consensus AP-1 binding site, TGACTGA (TCACTGA for SP- A2), which when mutated (TGAGAGT) prevented the formation of the TPA-induced DNA/protein complex. The TPA-induced complex was supershifted in the presence of antibody against the Fun family of proteins, but not the Fos family of proteins. These results suggest that the binding of AP-1 or an AP-1-like factor to the first intron of SP-A1 and SP-A2 may be involved in the phorbol ester inhibition of human SP-A gene expression.",
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Induction of AP-1 binding to intron 1 of SP-A1 and SP-A2 is implicated in the phorbol ester inhibition of human SP-A promoter activity. / Hoover, Russell R.; Pavlovic, Jelena; Floros, Joanna.

In: Experimental Lung Research, Vol. 26, No. 5, 17.07.2000, p. 303-317.

Research output: Contribution to journalArticle

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T1 - Induction of AP-1 binding to intron 1 of SP-A1 and SP-A2 is implicated in the phorbol ester inhibition of human SP-A promoter activity

AU - Hoover, Russell R.

AU - Pavlovic, Jelena

AU - Floros, Joanna

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N2 - A deletional analysis of the SP-A1 promoter in NCI-H441 cells was performed to identify potential cis-acting elements involved in phorbol ester-mediated repression of human SP-A transcription. The phorbol ester TPA reduced SP-A1 and SP-A2 promoter activity to approximately 35% to 45% compared to that of control cells. The inhibitory effect of TPA was significantly reduced upon removal of the region +64/+394 relative to the SP- A1 transcription start site. Using NCI-H441 nuclear proteins, electromobility shift assay analysis showed that the intron region +309/+329 of SP-A1 and the corresponding region of SP-A2 formed sequence-specific DNA/protein complexes that were induced by TPA exposure. The region +318/+324 of SP-A1 contains sequences similar to a consensus AP-1 binding site, TGACTGA (TCACTGA for SP- A2), which when mutated (TGAGAGT) prevented the formation of the TPA-induced DNA/protein complex. The TPA-induced complex was supershifted in the presence of antibody against the Fun family of proteins, but not the Fos family of proteins. These results suggest that the binding of AP-1 or an AP-1-like factor to the first intron of SP-A1 and SP-A2 may be involved in the phorbol ester inhibition of human SP-A gene expression.

AB - A deletional analysis of the SP-A1 promoter in NCI-H441 cells was performed to identify potential cis-acting elements involved in phorbol ester-mediated repression of human SP-A transcription. The phorbol ester TPA reduced SP-A1 and SP-A2 promoter activity to approximately 35% to 45% compared to that of control cells. The inhibitory effect of TPA was significantly reduced upon removal of the region +64/+394 relative to the SP- A1 transcription start site. Using NCI-H441 nuclear proteins, electromobility shift assay analysis showed that the intron region +309/+329 of SP-A1 and the corresponding region of SP-A2 formed sequence-specific DNA/protein complexes that were induced by TPA exposure. The region +318/+324 of SP-A1 contains sequences similar to a consensus AP-1 binding site, TGACTGA (TCACTGA for SP- A2), which when mutated (TGAGAGT) prevented the formation of the TPA-induced DNA/protein complex. The TPA-induced complex was supershifted in the presence of antibody against the Fun family of proteins, but not the Fos family of proteins. These results suggest that the binding of AP-1 or an AP-1-like factor to the first intron of SP-A1 and SP-A2 may be involved in the phorbol ester inhibition of human SP-A gene expression.

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