Type of Submission
Poster
Keywords
Tetrahymena, mitotic regulation, H3K27, acetylation, gene regulation, whole RNA sequencing
Proposal
The methylation and acetylation of histone tails is a critical part of gene regulation which has implications for homeostasis and disease. In cancers such as diffuse intrinsic pontine glioma (DIPG) where many patients are heterozygous for a mutation on histone H3 that replaces lysine with methionine, cells show a higher amount of acetylation and less trimethylation at this position. Tetrahymena thermophila are free-living, eukaryotic ciliates that are often used for epigenetic studies due to their rapid rate of cell division and the ease with which they may be cultured. We have previously shown that BAY-293, an inhibitor of the Ras-GEF Sos, significantly impacts mitotic rate in Tetrahymena thermophila. Since lysine 27 of histone H3 is apparently a critical residue impacting growth regulation, we asked the following questions: 1. Would treatment with BAY-293 impact H3K27ac in Tetrahymena thermophila as measured by immunofluorescence? 2. Would BAY-293 affect gene expression in this organism as measured by total RNA sequencing? Our results indicate that 90-minute exposure to BAY-293 significantly reduces acetylation at H3K27. Acetylation returns to normal once the drug has been washed out for 90 minutes. 90-minute exposure to BAY-293 also reduces a number of cellular RNA populations, including mRNA and rRNA.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.
Publication Date
2024
The Antimitotic Srug BAY-293 Reduces Acetylation at H3K27 and Decreases Diversity of Genes Expressed while Reducing Expression of rRNA and Ribosomal Proteins in Tetrahymena Thermophila
The methylation and acetylation of histone tails is a critical part of gene regulation which has implications for homeostasis and disease. In cancers such as diffuse intrinsic pontine glioma (DIPG) where many patients are heterozygous for a mutation on histone H3 that replaces lysine with methionine, cells show a higher amount of acetylation and less trimethylation at this position. Tetrahymena thermophila are free-living, eukaryotic ciliates that are often used for epigenetic studies due to their rapid rate of cell division and the ease with which they may be cultured. We have previously shown that BAY-293, an inhibitor of the Ras-GEF Sos, significantly impacts mitotic rate in Tetrahymena thermophila. Since lysine 27 of histone H3 is apparently a critical residue impacting growth regulation, we asked the following questions: 1. Would treatment with BAY-293 impact H3K27ac in Tetrahymena thermophila as measured by immunofluorescence? 2. Would BAY-293 affect gene expression in this organism as measured by total RNA sequencing? Our results indicate that 90-minute exposure to BAY-293 significantly reduces acetylation at H3K27. Acetylation returns to normal once the drug has been washed out for 90 minutes. 90-minute exposure to BAY-293 also reduces a number of cellular RNA populations, including mRNA and rRNA.
