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. 1992 Nov 11;20(21):5811–5817. doi: 10.1093/nar/20.21.5811

Purification and characterization of NF-R1 that regulates the expression of the human multidrug resistance (MDR1) gene.

M Ogura 1, T Takatori 1, T Tsuruo 1
PMCID: PMC334420  PMID: 1360648

Abstract

We have purified a protein, NF-R1, that specifically binds to two unrelated motifs--the ATTCAGTCA motif and the GC-box motif--on the MDR1 proximal promoter. Purified NF-R1 has been confirmed by southwestern blotting to be a 110-kDa protein. Methylation interference analysis revealed that the nucleotides were in close contact with purified NF-R1 on the ATTCAGTCA and GC-box motifs. The nucleotides were required for the binding of NF-R1, as seen by competition gel mobility shift assay using point mutated oligonucleotides. In a CAT expression assay using the corresponding point-mutated MDR1 promoter fused to a CAT gene, binding inhibition of NF-R1 to the promoter resulted in 2- to 3-fold increases of CAT activity, as compared to the intact promoter in Adriamycin-resistant K562 cells. Thus NF-R1 has a relation to the negative regulation of the MDR1 gene transcription.

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