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[Binding of transcription factors with functionally significant regions of MDR1 gene promoter during its activation]
L N Bozhenok1, A V Kolpakova, A V Vladimirova
1Institute of Bioorganic Chemistry, Siberian Branch of Russian Academy of Science, Novosibirsk.
Abstract:
Interaction of transcription factors with functional regions of gene mdr1 promoter has been studied. Binding of transcription factors to double-stranded (ds) oligonucleotides, mimicking four important regulatory regions before and after treatment of KB and K-562 cell lines with doxorubicin, cytarabine, and vinblastine was analysed by gel-shift assay. Gene induction resulted in enhancement of the main complex A(A1) proteins formation on the all four regions in both the cell lines. Inhibition of each complex formation by an excess of non-radiolabelled ds oligonucleotides suggests the possibility of regions interaction during promoter activation. Another indication of interaction of regulatory regions during activation is overlapping minor complexes sets. The results suggest that the four studied regulatory regions are important for mdr1 gene activation. Corresponding oligonucleotides mimicking these regions can be employed as inhibitors of gene transcription that bind specific transcriptional factors.
Insights
This study investigated transcription factor binding to the mdr1 gene promoter. Results show four key regulatory regions are crucial for mdr1 gene activation and can be targeted for transcription inhibition.
Area of Science:
- Molecular Biology
- Gene Regulation
Context:
- The mdr1 gene plays a role in multidrug resistance.
- Understanding its transcriptional regulation is key to developing new therapeutic strategies.
Purpose:
- To investigate the interaction of transcription factors with functional regions of the mdr1 gene promoter.
- To analyze the effect of chemotherapy drugs on transcription factor binding.
Summary:
- Gel-shift assays were used to study transcription factor binding to four regulatory regions of the mdr1 promoter in KB and K-562 cell lines before and after treatment with doxorubicin, cytarabine, and vinblastine.
- Gene induction enhanced the formation of main complex A(A1) proteins across all four regions in both cell lines.
- Inhibition studies and analysis of minor complexes suggest interaction between these regulatory regions during promoter activation.
Impact:
- The four studied regions are critical for mdr1 gene activation.
- Oligonucleotides mimicking these regions show potential as specific inhibitors of mdr1 gene transcription.