Aryl Hydrocarbon Receptor Ligands of Widely Different Toxic Equivalency Factors Induce Similar Histone Marks in Target Gene Chromatin

Posttranslational histone modifications are a critical regulatory mechanism of gene transcription. Previous studies from our laboratory have shown that contingent on binding to its cognate promoter motifs in the Cyp1a1 gene, activation of the aryl hydrocarbon receptor (AHR) by benzo[a]pyrene (BaP) treatment induces histone modifications in the Cyp1a1 promoter that are required for activation of gene transcription. Here, we have studied different AHR ligands, including polychlorinated biphenyls (PCBs) of different toxic equivalency factors (TEF), to determine whether changes in histone modifications are linked to different levels of Cyp1a1 expression or dependent on AHR-ligand affinity. We find that all ligands lead to the same pattern of histone modifications in a relationship that parallels the strength of their AHR-ligand affinity. Thus, whereas PCB126 (TEF 0.1), 3-methylcholanthrene, β-naphthoflavone, and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) initiate a pattern of histone marks similar to those induced by BaP, PCB77 (TEF 0.0001) causes a lower level of change in the same marks and requires a longer activation time than PCB126, BaP, or TCDD. In contrast, the non–dioxin-like PCB153 recruits AHR to the Cyp1a1 enhancer causing a displacement of enhancer-associated histone H3 but does not cause the other observed histone mark changes nor does it induce transcription. These results indicate that AHR recruitment to the promoter is not sufficient to induce the histone modifications needed to activate gene expression and show that there is a good correlation between the regulatory chromatin changes associated with ligand-induced AHR target gene transcription and the resultant toxicity of the ligand

Airborne exposures associated with the typical use of an aerosol brake cleaner during vehicle repair work

<p>Many petroleum-based products are used for degreasing and cleaning purposes during vehicle maintenance and repairs. Although prior studies have evaluated chemical exposures associated with this type of work, most of these have focused on gasoline and exhaust emissions, with few samples collected solely during the use of an aerosol cleaning product. In this case study, we assess the type of airborne exposures that would be expected from the typical use of an aerosol brake cleaner during vehicle repair work. Eight exposure scenarios were evaluated over a 2-day study in which the benzene content of the brake cleaner and potential for dilution ventilation and air flow varied. Both short-term (15 min) and task-based (≥1 hr) charcoal tube samples were collected in the breathing zone and adjacent work area and analyzed for total hydrocarbons (THCs), toluene, and benzene. The majority of personal (N = 48) and area (N = 47) samples had detectable levels of THC and toluene, but no detections of benzene were found. For the personal short-term samples, average airborne concentrations ranged from 3.1–61.5 ppm (13.8–217.5 mg/m³) for THC and 2.2–44.0 ppm (8.2–162.5 mg/m³) for toluene, depending on the scenario. Compared to the personal short-term samples, average concentrations were generally 2–3 times lower for the personal task-based samples and 2–5 times lower for the area short-term samples. The highest exposures occurred when the garage bay doors were closed, floor fan was turned off, or greatest amount of brake cleaner was used. These findings add to the limited dataset on this topic and can be used to bound or approximate worker or consumer exposures from use of aerosol cleaning products with similar compositions and use patterns.</p

Formaldehyde-Assisted Isolation of Regulatory Elements (FAIRE) analysis uncovers broad changes in chromatin structure resulting from hexavalent chromium exposure.

The ability of chromatin to switch back and forth from open euchromatin to closed heterochromatin is vital for transcriptional regulation and genomic stability, but its dynamic structure is subject to disruption by exposure to environmental agents such as hexavalent chromium. Cr(VI) exposure disrupts chromatin remodeling mechanisms and causes chromosomal damage through formation of free radicals, Cr-DNA adducts, and DNA-Cr-protein cross-links. In addition, acute, high-concentration, and chronic, low-concentration exposures to Cr(VI) lead to significantly different transcriptional and genomic stability outcomes. We used mouse hepatoma Hepa-1c1c7 cells to investigate how transcriptional responses to chromium treatment might correlate with structural chromatin changes. We used Formaldehyde-Assisted Isolation of Regulatory Elements (FAIRE) analysis coupled with deep sequencing to identify regions of the genome that may switch between open and closed chromatin in response to exposure to varying Cr(VI) concentrations. At either Cr(VI) concentration, chromatin domains surrounding binding sites for AP-1 transcription factors become significantly open, whereas BACH2 and CTCF binding sites are open solely at the low and high concentrations, respectively. Parallel gene expression profiling using RNA-seq indicates that the structural chromatin changes caused by Cr(VI) affect gene expression levels in the target areas that vary depending on Cr(VI) concentration, but show no correlation between global changes in the overall transcriptional response and Cr(VI) concentration. Our results suggest that FAIRE may be a useful technique to map chromatin elements targeted by DNA damaging agents for which there is no prior knowledge of their specificity, and to identify subsequent transcriptomic changes induced by those agents

Enrichment of Genomic Elements in FAIRE Peaks Opened by Cr(VI) Treatment.

<p>Enrichment of Genomic Elements in FAIRE Peaks Opened by Cr(VI) Treatment.</p

Opening of FAIRE peaks in promoter regions by chromium treatment is significantly correlated with changes in gene expression.

<p>(<b>A</b>) Venn diagram of concordant (<i>p</i> = 3.75×10⁻⁹⁹) gene expression changes induced by long-term low dose and acute high dose Cr(VI) treatments. (<b>B</b>) Hierarchical clustering of the 442 genes with concordant expression. (<b>C</b>) gene expression changes corresponding to peaks opened by Cr(VI) in promoter regions within 1 kb of the TSS. In the low concentration, long term Cr(VI) treatment, 199 of the 252 FAIRE peaks were contained in up-regulated genes (<i>p</i> = 2.79×10⁻¹⁶) and 202 of 243 peaks in the acute, high concentration treatment were contained in down-regulated genes (<i>p</i> = 4.98×10⁻¹⁰).</p