Title:

Assessing the Histone Methyltransferase Activity of SET Domain of AP2VIIa-7 in Toxoplasma gondii

Toxoplasma gondii is an apicomplexan parasite that relies on tight gene regulation to switch between life stages. The BDP complex, an essential transcription regulator, contains three bromodomain proteins (BDPs), BDP1, BDP2, and BDP5, and a SET methyltransferase, AP2VIIa-7. Inducible knockdown of BDP1 led to significant gene dysregulation, including the upregulation of several sexual stage genes. The repression of sexual stage genes during the asexual tachyzoite stage, which we study, is mediated, in part, by methylation. We predict that AP2VIIa-7 is a key histone methyltransferase and that histone methylation mediated by AP2VIIa-7 is inhibited when not properly recruited to chromatin during BDP1 loss. To address this, histone methylation activity of a recombinant AP2VIIa-7 SET domain will be measured via a histone methylation assay, and changes in global histone methylation levels during AP2VIIa-7 knockdown will be assessed via western blotting. This approach will validate the SET domain's enzymatic activity as a methyltransferase and identify the specific histone residues that the SET domain in AP2VIIa-7 methylates. This will allow further understanding of sexual stage genes and aid in finding ways to control the transmission of T. gondii through targeting and preventing sexual reproduction.

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