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Butylated hydroxytoluene modulates DNA methylation in rats
B F Vanyushin1, N G Lopatina, C K Wise
1Division of Molecular Basis of Ontogenesis, A.N. Belozersky Institute of Physico-Chemical Biology, M.V. Lomonosov Moscow State University, Russia. Vanyushin@moo.genebee.msu.su
European Journal of Biochemistry
|October 21, 1998
Summary
Butylated hydroxytoluene (BHT) significantly increases DNA methyltransferase activity in rat organs within hours, promoting de novo DNA methylation. These BHT-induced changes in methylation are tissue-specific and reversible.
Area of Science:
- Biochemistry
- Molecular Biology
- Toxicology
Background:
- DNA methylation is crucial for gene regulation and cellular function.
- Butylated hydroxytoluene (BHT) is a synthetic antioxidant with known biological effects.
- Understanding BHT's impact on DNA methylation machinery is important for assessing its toxicological profile.
Purpose of the Study:
- To investigate the effect of a single intraperitoneal injection of BHT on DNA methyltransferase activity in various rat organs.
- To characterize the changes in DNA methyltransferase activity and DNA methylation patterns induced by BHT.
- To explore the impact of BHT on specific genes and their methylation status.
Main Methods:
- Intraperitoneal injection of BHT (60 mg/kg) in male rats.
- Assay of nuclear DNA(cytosine-5)-methyl transferase activity in liver, kidneys, heart, spleen, brain, and lungs.
- Isoelectric focusing electrophoresis to analyze methyltransferase protein fractions.
- Analysis of S-adenosylmethionine and S-adenosylhomocysteine levels.
- Assessment of 5-methyldeoxycytidine content in DNA.
- Gene-specific methylation analysis (cytosine DNA-methyl transferase gene, c-Ha-ras gene).
- Measurement of mRNA transcripts for S-adenosylmethionine synthetase and p53.
Main Results:
- BHT injection caused a rapid and significant increase in methyltransferase activity across multiple organs, peaking at 8 hours.
- BHT altered the profile of methyltransferase protein fractions, with induced enzymes preferentially methylating unmethylated DNA (de novo methylation).
- BHT induced tissue-specific changes in DNA methylation, including hypomethylation of specific genes (renal methyl transferase, hepatic c-Ha-ras), and altered 5-methyldeoxycytidine content in some organs.
- BHT increased hepatic mRNA for S-adenosylmethionine synthetase and p53 but did not affect methyltransferase homologous mRNA.
Conclusions:
- BHT administration induces a robust, transient increase in DNA methyltransferase activity in various rat organs.
- The induced methyltransferase exhibits a preference for de novo DNA methylation.
- BHT causes reversible, tissue-specific alterations in DNA methylation patterns and gene methylation status.
- BHT's effects on DNA methylation may contribute to its overall biological and toxicological actions.