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Cellular Oncology
Published in: Cellular Oncology 5/2016

01-10-2016 | Original Paper

MGMT DNA repair gene promoter/enhancer haplotypes alter transcription factor binding and gene expression

Authors: Meixiang Xu, Courtney E. Cross, Jordan T. Speidel, Sherif Z. Abdel-Rahman

Published in: Cellular Oncology | Issue 5/2016

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Abstract

Background

The O6-methylguanine-DNA methyltransferase (MGMT) protein removes O6-alkyl-guanine adducts from DNA. MGMT expression can thus alter the sensitivity of cells and tissues to environmental and chemotherapeutic alkylating agents. Previously, we defined the haplotype structure encompassing single nucleotide polymorphisms (SNPs) in the MGMT promoter/enhancer (P/E) region and found that haplotypes, rather than individual SNPs, alter MGMT promoter activity. The exact mechanism(s) by which these haplotypes exert their effect on MGMT promoter activity is currently unknown, but we noted that many of the SNPs comprising the MGMT P/E haplotypes are located within or in close proximity to putative transcription factor binding sites. Thus, these haplotypes could potentially affect transcription factor binding and, subsequently, alter MGMT promoter activity.

Methods

In this study, we test the hypothesis that MGMT P/E haplotypes affect MGMT promoter activity by altering transcription factor (TF) binding to the P/E region. We used a promoter binding TF profiling array and a reporter assay to evaluate the effect of different P/E haplotypes on TF binding and MGMT expression, respectively.

Results

Our data revealed a significant difference in TF binding profiles between the different haplotypes evaluated. We identified TFs that consistently showed significant haplotype-dependent binding alterations (p ≤ 0.01) and revealed their role in regulating MGMT expression using siRNAs and a dual-luciferase reporter assay system.

Conclusions

The data generated support our hypothesis that promoter haplotypes alter the binding of TFs to the MGMT P/E and, subsequently, affect their regulatory function on MGMT promoter activity and expression level.
Appendix
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Metadata
Title
MGMT DNA repair gene promoter/enhancer haplotypes alter transcription factor binding and gene expression
Authors
Meixiang Xu
Courtney E. Cross
Jordan T. Speidel
Sherif Z. Abdel-Rahman
Publication date
01-10-2016
Publisher
Springer Netherlands
Published in
Cellular Oncology / Issue 5/2016
Print ISSN: 2211-3428
Electronic ISSN: 2211-3436
DOI
https://doi.org/10.1007/s13402-016-0286-4

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