Identification of two nuclear protein binding sites and their role in the regulation of the murine multidrug resistance mdr1a promoter

D. Cohen, L. Yu, R. Rzepka, Susan Band Horwitz

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Multidrug resistance genes (mdr) that encode P-glycoproteins (P-gp) are transcriptionally regulated in normal tissues and in some multidrug- resistant (MDR) cells. Several lines of evidence suggest that regulation of P-gp overexpression at the transcriptional level is also important in human tumors. In murine MDR cells, mdr1a and/or mdr1b genes are overexpressed and P-gp isoforms are overproduced. To identify the mdr1a promoter regions that are required for transcription, the promoter has been linked to the chloramphenicol acetyltransferase (CAT) gene in transient expression vectors. 5'-Deletions of the promoter sequences have demonstrated that the region between -155 to +89 bp is crucial for basal activity of the mdr1a gene. DNase I footprinting, methylation interference, and gel retardation assays identified two nuclear protein binding sites within these sequences. One of the nuclear protein binding sites contains an 11-bp DNA sequence that interacts with nuclear protein(s) and is conserved in the promoters of the murine mdr1a and mdr1b, hamster pgp1, and human MDR1 genes. The conserved SP1 site (5'-GGGCGGG-3') that is present further downstream was shown to interact with its nuclear factor. These observations suggest that at least part of mdr gene transcriptional regulation is mediated by conserved mdr cis-regulatory elements and common nuclear factors.

Original languageEnglish (US)
Pages (from-to)641-649
Number of pages9
JournalDNA and Cell Biology
Volume13
Issue number6
StatePublished - 1994
Externally publishedYes

Fingerprint

Multiple Drug Resistance
Nuclear Proteins
Protein Binding
P-Glycoproteins
MDR Genes
Binding Sites
Genes
Chloramphenicol O-Acetyltransferase
Sequence Deletion
Deoxyribonuclease I
Electrophoretic Mobility Shift Assay
Genetic Promoter Regions
Cricetinae
Methylation
Protein Isoforms
Neoplasms

ASJC Scopus subject areas

  • Cell Biology
  • Genetics
  • Molecular Biology

Cite this

Identification of two nuclear protein binding sites and their role in the regulation of the murine multidrug resistance mdr1a promoter. / Cohen, D.; Yu, L.; Rzepka, R.; Band Horwitz, Susan.

In: DNA and Cell Biology, Vol. 13, No. 6, 1994, p. 641-649.

Research output: Contribution to journalArticle

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