The exploration of N6-deoxyadenosine methylation in mammalian genomes.

N6-methyladenine (N6-mA, m6dA, or 6mA), a prevalent DNA modification in prokaryotes, has recently been identified in higher eukaryotes, including mammals. Although 6mA has been well-studied in prokaryotes, the function and regulatory mechanism of 6mA in eukaryotes are still poorly understood. Recent studies indicate that 6mA can serve as an epigenetic mark and play critical roles in various biological processes, from transposable-element suppression to environmental stress response. Here, we review the significant advances in methodology for 6mA detection and major progress in understanding the regulation and function of this non-canonical DNA methylation in eukaryotes, predominantly mammals.

Effect of atrazine and chlorpyrifos exposure on cytochrome P450 contents and enzyme activities in common carp gills.

Chlorpyrifos (CPF) and atrazine (ATR) are the most widely used organophosphate insecticides and triazine herbicides, respectively, worldwide. This study aimed at investigating the effects of ATR, CPF and mixture on common carp gills following 40-d exposure and 40-d recovery experiments. Cytochrome P450 content, activities of aminopyrine N-demethylase (APND) and erythromycin N-demethylase (ERND) and the mRNA levels of the CYP1 family (CYP1A, CYP1B, and CYP1C) were determined. In total, 220 common carps were divided into eleven groups, and each group was treated with a specific concentration of ATR (4.28, 42.8 and 428 µg/L), CPF (1.16, 11.6 and 116 µg/L) or ATR-CPF mixture (1.13, 11.3 and 113 µg/L). The results showed that P450 content and activities of APND and ERND in fish exposed to ATR and mixture were significantly higher than those in the control group. After the 40-d recovery treatment (i.e., depuration), the P450 content and the activities of APND and ERND in fish decreased to the background levels. A similar tendency was also found in the mRNA levels of the CYP1 family (CYP1A, CYP1B, and CYP1C) in common carp gills. The CPF-treated fish showed no significant difference from the control groups, except for a significant CYP1C induction. These results indicated that CYP enzyme levels are induced by ATR but were only slightly affected by CPF in common carp gills. In addition, the ATR and CPF exposure showed an antagonistic effect on P450 enzymes in common carp gills.

Induction of cytochrome P450 isozymes in human amnion FL cells.

The human epithelial FL cell line contains low levels of constitutive ethoxyresorufin O-deethylase, ethoxycoumarin O-deethylase and aminopyrine N-demethylase activities, of which ethoxyresorufin O-deethylase could be induced by norepinephrine, beta-naphthoflavone and 3-methylcholanthrene, but ethoxy-couarin O-deethylase and aminopyrine N-demethylase activities could be induced by norepinephrine, beta-naphthoflavone and 3-methylcholanthrene as well as phenobarbital. Inducibility of ethoxyresorufin O-deethylase, ethoxycoumarin O-deethylase and aminopyrine N-demethylase activities was 3.1-6.7-, 1.8-3.1- and 1.4-2.0-fold respectively. Co-treatment with 3-methylcholanthrene and norepinephrine resulted in higher ethoxyresorufin O-deethylase and aminopyrine N-demethylase activities than treatment with 3-methylcholanthrene alone. The induced ethoxyresorufin O-deethylase and ethoxycoumarin O-deethylase activities remained at a high level for 24-36 h after removal of the inducer from the medium. The metabolizing ability of induced FL cells is stronger than that of non-induced ones in a given time following induction. The results further confirm that it is feasible to use the cytochrome P450 isozyme-induced FL cell as a biological indicator in short-term tests for screening promutagens/procarcinogens.