Analysis of the tissue-specific promoter of the MUC1 gene

Ales Kovarik, Nigel Peat, David Wilson, Sandra J Gendler, Joyce Taylor-Papadimitriou

Research output: Contribution to journalArticle

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Abstract

The 5′-sequences flanking the human MUC1 gene have been analyzed for their ability to direct expression of a reporter gene (the chloramphenicol acetyltransferase gene (CAT)) in cell lines that normally express or do not express the MUC1 gene. A construct containing 2.9 kilobase pairs of MUC1 5′-flanking sequence sequence showed expression of CAT in breast and pancreatic cell lines but not in the non-epithelial cell lines HT-1080, SK23, and HTB96. Deletion analysis showed that maximum expression was obtained in ZR-75 (breast cancer line) and HPAF (pancreatic cancer line) with only 743 base pairs of 5′-flanking sequence. Sequences within 1.6 kilobase pairs of the transcriptional start site showed enhancing activity in a vector carrying an enhancerless SV40 promoter. Analysis of proximal 5′-sequences in a promoterless CAT vector carrying the SV40 enhancer showed that sequences between -60 and -150 were crucial for tissue-specific expression. An Sp1 site at -99/-90 and an E box (E-MUC1) at -84/-64 in this region were shown by mutational analysis to play a role in the regulation of transcription. Gel shift analysis with oligonucleotides and nuclear extracts of ZR-75 showed protein binding to both of these sites. Sp1 binding activity was similar in ZR-75 and HT1080 cells, whereas binding of factors to the E-MUC1 oligonucleotide revealed quantitative and qualitative differences between epithelial and non-epithelial cells.

Original languageEnglish (US)
Pages (from-to)9917-9926
Number of pages10
JournalJournal of Biological Chemistry
Volume268
Issue number13
StatePublished - May 5 1993
Externally publishedYes

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Genes
Chloramphenicol O-Acetyltransferase
Tissue
5' Flanking Region
Cells
Oligonucleotides
Cell Line
Electrophoretic Mobility Shift Assay
Pancreatic Neoplasms
Reporter Genes
Protein Binding
Base Pairing
Transcription
Breast
Breast Neoplasms
Gels

ASJC Scopus subject areas

  • Biochemistry

Cite this

Kovarik, A., Peat, N., Wilson, D., Gendler, S. J., & Taylor-Papadimitriou, J. (1993). Analysis of the tissue-specific promoter of the MUC1 gene. Journal of Biological Chemistry, 268(13), 9917-9926.

Analysis of the tissue-specific promoter of the MUC1 gene. / Kovarik, Ales; Peat, Nigel; Wilson, David; Gendler, Sandra J; Taylor-Papadimitriou, Joyce.

In: Journal of Biological Chemistry, Vol. 268, No. 13, 05.05.1993, p. 9917-9926.

Research output: Contribution to journalArticle

Kovarik, A, Peat, N, Wilson, D, Gendler, SJ & Taylor-Papadimitriou, J 1993, 'Analysis of the tissue-specific promoter of the MUC1 gene', Journal of Biological Chemistry, vol. 268, no. 13, pp. 9917-9926.
Kovarik A, Peat N, Wilson D, Gendler SJ, Taylor-Papadimitriou J. Analysis of the tissue-specific promoter of the MUC1 gene. Journal of Biological Chemistry. 1993 May 5;268(13):9917-9926.
Kovarik, Ales ; Peat, Nigel ; Wilson, David ; Gendler, Sandra J ; Taylor-Papadimitriou, Joyce. / Analysis of the tissue-specific promoter of the MUC1 gene. In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 13. pp. 9917-9926.
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