Neutrophil elastase stimulates MUC1 gene expression through increased Sp1 binding to the MUC1 promoter

Ippei Kuwahara, Erik P. Lillehoj, Akinori Hisatsune, Wenju Lu, Yoichiro Isohama, Takeshi Miyata, K. Chul Kim

Research output: Contribution to journalArticle

Abstract

We previously reported MUC1 was a cell surface receptor for Pseudomonas aeruginosa, and binding of bacteria to cells was significantly reduced by pretreatment with neutrophil elastase (NE) (Lillehoj EP, Hyun SW, Kim BT, Zhang XG, Lee DI, Rowland S, and Kim KC. Am J Physiol Lung Cell Mol Physiol 280: L181-L187, 2001). The current study was conducted to ascertain NE effects on MUC1 gene transcription, and MUC1 protein synthesis and degradation. A549 human lung carcinoma cells treated with NE exhibited significantly higher MUC1 protein levels in detergent lysates compared with cells treated with vehicle alone. Also, MUC1 protein shed into cell-conditioned medium was rapidly and completely degraded by NE. Actinomycin D blocked NE-stimulated increase in MUC1 protein expression, suggesting a mechanism of increased gene transcription that was confirmed by measurement of quantitatively greater MUC1 mRNA levels in NE-treated cells compared with controls. However, NE did not alter MUC1 mRNA stability, implying increased de novo transcription induced by the protease. NE increased promoter activity in A549 cells transfected with MUC1 gene promoter-luciferase reporter plasmid. This effect of NE was completely blocked by mithramycin A, an inhibitor of Sp1, as well as mutation of one of the putative Sp1 binding sites in MUC1 promoter located at -99/-90 relative to transcription initiation site. EMSA revealed NE enhanced binding of Sp1 to this 10-bp segment in a time-dependent manner. These results indicate the increase in MUC1 gene transcription by NE is mediated through increase in Sp1 binding to -99/-90 segment of MUC1 promoter.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume289
Issue number2 33-2
DOIs
StatePublished - Aug 2005
Externally publishedYes

Fingerprint

Leukocyte Elastase
Gene Expression
Genes
Lung
Proteins
Transcription Initiation Site
RNA Stability
Cell Surface Receptors
Dactinomycin
Conditioned Culture Medium
Luciferases
Detergents
Pseudomonas aeruginosa
Proteolysis
Plasmids
Peptide Hydrolases
Binding Sites

Keywords

  • Epithelial
  • Mithramycin A
  • Mucin
  • Protease
  • Transcription

ASJC Scopus subject areas

  • Pulmonary and Respiratory Medicine
  • Cell Biology
  • Physiology

Cite this

Neutrophil elastase stimulates MUC1 gene expression through increased Sp1 binding to the MUC1 promoter. / Kuwahara, Ippei; Lillehoj, Erik P.; Hisatsune, Akinori; Lu, Wenju; Isohama, Yoichiro; Miyata, Takeshi; Kim, K. Chul.

In: American Journal of Physiology - Lung Cellular and Molecular Physiology, Vol. 289, No. 2 33-2, 08.2005.

Research output: Contribution to journalArticle

Kuwahara, Ippei ; Lillehoj, Erik P. ; Hisatsune, Akinori ; Lu, Wenju ; Isohama, Yoichiro ; Miyata, Takeshi ; Kim, K. Chul. / Neutrophil elastase stimulates MUC1 gene expression through increased Sp1 binding to the MUC1 promoter. In: American Journal of Physiology - Lung Cellular and Molecular Physiology. 2005 ; Vol. 289, No. 2 33-2.
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