cis-Acting elements that confer lung epithelial cell expression of the CC10 gene

B. R. Stripp, P. L. Sawaya, D. S. Luse, K. A. Wikenheiser, S. E. Wert, J. A. Huffman, D. L. Lattier, Gurmukh Singh, S. L. Katyal, J. A. Whitsett

Research output: Contribution to journalArticle

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Abstract

To define cis-acting genetic elements responsible for cell-specific transcriptional regulation of the CC10 gene, DNA sequences spanning nucleotides -2338 to +49 of the rat CC10 gene were linked to a reporter gene coding for chloramphenicol acetyltransferase (CAT). In transient expression assays, CC10 sequences were capable of restricting CAT expression to a human lung adenocarcinoma cell line similar to pulmonary Clara cells. Transgenic mice harboring the hybrid RtCC10-CAT construct expressed high levels of CAT activity specifically within protein extracts of lung and trachea. Transcripts for the CAT reporter gene colocalized with those for the endogenous murine CC10 gene within the airways of transgenic mice. Functional analysis of deletion mutants identified stimulatory, inhibitory, and cell type-specific transcriptional regulatory elements. The results of gel retention and DNaseI protection assays suggest that a transcriptional stimulatory region located between -320 and -175, and a cell type-specific regulatory element located between -175 and +49, result from a series of protein-DNA interactions occurring at -220 to -205 and -128 to -86, respectively. Lung epithelial specific transcriptional regulatory elements described herein will be useful for expression of chimeric genes within epithelial cells lining the trachea, bronchi, and bronchioles of mice.

Original languageEnglish (US)
Pages (from-to)14703-14712
Number of pages10
JournalJournal of Biological Chemistry
Volume267
Issue number21
StatePublished - Jan 1 1992
Externally publishedYes

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Chloramphenicol O-Acetyltransferase
Genes
Epithelial Cells
Gene Expression
Lung
Transcriptional Regulatory Elements
Trachea
Reporter Genes
Transgenic Mice
Assays
Bronchioles
Bronchi
Functional analysis
DNA sequences
Linings
Proteins
Nucleotides
Gels
Rats
Cell Line

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Stripp, B. R., Sawaya, P. L., Luse, D. S., Wikenheiser, K. A., Wert, S. E., Huffman, J. A., ... Whitsett, J. A. (1992). cis-Acting elements that confer lung epithelial cell expression of the CC10 gene. Journal of Biological Chemistry, 267(21), 14703-14712.

cis-Acting elements that confer lung epithelial cell expression of the CC10 gene. / Stripp, B. R.; Sawaya, P. L.; Luse, D. S.; Wikenheiser, K. A.; Wert, S. E.; Huffman, J. A.; Lattier, D. L.; Singh, Gurmukh; Katyal, S. L.; Whitsett, J. A.

In: Journal of Biological Chemistry, Vol. 267, No. 21, 01.01.1992, p. 14703-14712.

Research output: Contribution to journalArticle

Stripp, BR, Sawaya, PL, Luse, DS, Wikenheiser, KA, Wert, SE, Huffman, JA, Lattier, DL, Singh, G, Katyal, SL & Whitsett, JA 1992, 'cis-Acting elements that confer lung epithelial cell expression of the CC10 gene', Journal of Biological Chemistry, vol. 267, no. 21, pp. 14703-14712.
Stripp BR, Sawaya PL, Luse DS, Wikenheiser KA, Wert SE, Huffman JA et al. cis-Acting elements that confer lung epithelial cell expression of the CC10 gene. Journal of Biological Chemistry. 1992 Jan 1;267(21):14703-14712.
Stripp, B. R. ; Sawaya, P. L. ; Luse, D. S. ; Wikenheiser, K. A. ; Wert, S. E. ; Huffman, J. A. ; Lattier, D. L. ; Singh, Gurmukh ; Katyal, S. L. ; Whitsett, J. A. / cis-Acting elements that confer lung epithelial cell expression of the CC10 gene. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 21. pp. 14703-14712.
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