Sp3 regulates Fas expression in lung epithelial cells

Huiling Pang, Kathleen Miranda, Alan Fine

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

By transducing an apoptotic signal in immune effector cells, Fas has been directly implicated in the control of immunological activity. Expression and functional results, however, have also suggested a role for Fas in regulating cell turnover in specific epithelial populations. To characterize factors responsible for Fas expression in epithelial cells, approximately 3 kb of the 5@? flanking region of the mouse Fas gene was isolated. By rapid amplification of cDNA ends and primer extension, transcriptional start sites were identified within 50 bp upstream of the translation start site. Transient transfection of promoter-luciferase constructs in a mouse lung epithelial cell line, MLE-15, localized promoter activity to the first 77 bp of upstream sequence. By using a 60bp DNA probe (-18 to -77) in electrophoretic mobility-shift assays, three shifted complexes were found. Incubation with excess cold Sp1 oligonucleotide or an anti-Sp3 antibody inhibited complex formation. Site-directed mutagenesis of the Sp1 site resulted in 60-70% loss of promoter activity. In Drosophila SL-2 cells, promoter activity was markedly increased by co-transfection of an Sp3 expression construct. These results show that the Sp3 protein is involved in regulating Fas gene expression in lung epithelial cells.

Original languageEnglish (US)
Pages (from-to)209-213
Number of pages5
JournalBiochemical Journal
Volume333
Issue number1
DOIs
StatePublished - Jul 1 1998

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Epithelial Cells
Lung
Transfection
Electrophoretic mobility
Mutagenesis
Maximum likelihood estimation
DNA Probes
Electrophoretic Mobility Shift Assay
Site-Directed Mutagenesis
Luciferases
Gene expression
Oligonucleotides
Drosophila
Amplification
Anti-Idiotypic Antibodies
Assays
Complementary DNA
Genes
Gene Expression
Cell Line

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sp3 regulates Fas expression in lung epithelial cells. / Pang, Huiling; Miranda, Kathleen; Fine, Alan.

In: Biochemical Journal, Vol. 333, No. 1, 01.07.1998, p. 209-213.

Research output: Contribution to journalArticle

Pang, Huiling ; Miranda, Kathleen ; Fine, Alan. / Sp3 regulates Fas expression in lung epithelial cells. In: Biochemical Journal. 1998 ; Vol. 333, No. 1. pp. 209-213.
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