Tissue-specific enhancer of the human multidrug-resistance (MDR1) gene

K. Kohno, S. Sato, T. Uchiumi, H. Takano, S. Kato, M. Kuwano

Research output: Contribution to journalArticle

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Abstract

Identification of cis-regulatory sequences is a first step in analyzing the regulation of the human multidrug-resistant 1 (MDR1) gene which encodes the 170-kilodalton membrane P-glycoprotein in normal tissues and tumor cells. We have studied several overlapping genomic clones containing the 5'-flanking region of the gene. These clones span about 30 kilobases (kb) of contiguous DNA containing 10 kb of the gene and 20 kb of the 5'-flanking sequence. The nucleotide sequence of the first exon and the 2 kb preceding the exon were determined. DNA sequences containing the 5'-flanking regions were linked to the chloramphenicol acetyltransferase (CAT) gene. For transient CAT assay, we have employed six cell lines, including human cancer KB, vincristine-resistant VJ-300 derived from KB, mouse adrenal tumor Y-1, African green monkey kidney CV-1, mouse fibroblast NIH3T3, and human adrenal carcinoma SW-13 cells. Promoter activity was very weak regardless of the length of the promoter region in mouse adrenal tumor Y-1 and monkey kidney CV-1 cells, in which endogenous P-glycoprotein was expressed. Introduction of a 700-base genomic DNA fragment from a site located at 10 kb far upstream of the initiation site increased the transcription of the CAT gene in Y-1, CV-1, and SW-13 cells. However, no significant increase in the CAT activity could be observed in NIH3T3, KB, and VJ-300 cells. This fragment markedly augmented the expression of the CAT gene regardless of orientation or position, and it acted in a cell type-specific manner even with heterogeneous promoters. Our present study suggests that the 700-base pair fragment may carry a tissue-specific transcriptional enhancer that is active in at least some adrenal and kidney-derived cell lines.

Original languageEnglish
Pages (from-to)19690-19696
Number of pages7
JournalJournal of Biological Chemistry
Volume265
Issue number32
Publication statusPublished - Dec 17 1990
Externally publishedYes

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Multiple Drug Resistance
Chloramphenicol O-Acetyltransferase
Genes
Tissue
5' Flanking Region
Tumors
Glandular and Epithelial Neoplasms
Cells
P-Glycoprotein
Kidney
Exons
Clone Cells
Cercopithecus aethiops
Cell Line
Transcription Initiation Site
DNA sequences
DNA
Vincristine
Transcription
Fibroblasts

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Kohno, K., Sato, S., Uchiumi, T., Takano, H., Kato, S., & Kuwano, M. (1990). Tissue-specific enhancer of the human multidrug-resistance (MDR1) gene. Journal of Biological Chemistry, 265(32), 19690-19696.

Tissue-specific enhancer of the human multidrug-resistance (MDR1) gene. / Kohno, K.; Sato, S.; Uchiumi, T.; Takano, H.; Kato, S.; Kuwano, M.

In: Journal of Biological Chemistry, Vol. 265, No. 32, 17.12.1990, p. 19690-19696.

Research output: Contribution to journalArticle

Kohno, K, Sato, S, Uchiumi, T, Takano, H, Kato, S & Kuwano, M 1990, 'Tissue-specific enhancer of the human multidrug-resistance (MDR1) gene', Journal of Biological Chemistry, vol. 265, no. 32, pp. 19690-19696.
Kohno K, Sato S, Uchiumi T, Takano H, Kato S, Kuwano M. Tissue-specific enhancer of the human multidrug-resistance (MDR1) gene. Journal of Biological Chemistry. 1990 Dec 17;265(32):19690-19696.
Kohno, K. ; Sato, S. ; Uchiumi, T. ; Takano, H. ; Kato, S. ; Kuwano, M. / Tissue-specific enhancer of the human multidrug-resistance (MDR1) gene. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 32. pp. 19690-19696.
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