A Y-box consensus sequence is required for basal expression of the human multidrug resistance (mdr1) gene

M. E. Goldsmith, M. J. Madden, C. S. Morrow, K. H. Cowan

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Abstract

Basal transcription of the human multidrug resistance (mdr1) promoter was studied by chloramphenicol acetyltransferase (CAT) reporter fusion gene analysis in two parental and doxorubicin-resistant human tumor cell lines. Deletion of mdr1 DNA sequences to -89 relative to the start of transcription (at +1) had little effect on expression. Deletion of nucleotide sequences from -89 to -70, however, resulted in a 5-10-fold reduction in mdrCAT expression. DNase I footprint analysis demonstrated that the region from -85 to -70 was protected from nuclease digestion using nuclear extracts from these cell lines. The sequence between -82 and -73 is perfectly homologous with the 10-base pair Y-box consensus sequence found in the promoters of all major histocompatibility complex class-II (MHC II) genes. The Y-box sequence in MHC II genes is required for accurate and efficient transcription and contains the sequence CCAAT in the reverse orientation (Dorn, A., Durand, B., Marfing, C., Le Meur, M., Benoist, C., and Mathis, D. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 6249-6253). Mutations in the reverse CCAAT sequence of the Y-box consensus substantially reduced expression of an mdrCAT vector and eliminated nucleoprotein binding in an electrophoretic mobility shift assay. These results suggest that proteins which bind to the putative Y-box consensus sequence are critical for basal transcriptional regulation of the human mdr1 gene.

Original languageEnglish (US)
Pages (from-to)5856-5860
Number of pages5
JournalJournal of Biological Chemistry
Volume268
Issue number8
StatePublished - Jan 1 1993

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MDR Genes
Consensus Sequence
MHC Class II Genes
Transcription
Genes
Major Histocompatibility Complex
Chloramphenicol O-Acetyltransferase
Nucleoproteins
Deoxyribonuclease I
Cells
Multiple Drug Resistance
Electrophoretic Mobility Shift Assay
Tumor Cell Line
Reporter Genes
Base Pairing
Electrophoretic mobility
Doxorubicin
Digestion
DNA sequences
Tumors

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A Y-box consensus sequence is required for basal expression of the human multidrug resistance (mdr1) gene. / Goldsmith, M. E.; Madden, M. J.; Morrow, C. S.; Cowan, K. H.

In: Journal of Biological Chemistry, Vol. 268, No. 8, 01.01.1993, p. 5856-5860.

Research output: Contribution to journalArticle

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