Category
Poster - Basic
Description
Histone acetyltransferases (HATs) are enzymes involved in the transcriptional activation of genes. p300, a known HAT enzyme, acetylates histone tails at lysine and arginine residues. When histones are acetylated, they are more prone to promote transcriptional activation. Therefore, investigation into different HATs can aid in understanding gene regulation. Within C. neoformans, a pathogenic strain of yeast, there are a couple of putative HAT enzymes that are the focus of our research, the main one being the SAS3 gene, which shows a significant upregulation in expression when C. neoformans is allowed to grow in the absence of glucose . However, although the sequences of these genes suggest they are HAT enzymes, there is no definitive proof existing about their enzymatic activity. Our research aims to first isolate these proteins that have been cloned into a bacterial vector (pet45b(+)) via a His-tag purification method. Once isolated, the proteins can be assessed for their potential HAT activity via HAT assay. Every HAT assay requires an Acetyl-CoA source, substrate, and enzyme. Our current working HAT assay protocol involves using core histones purified from HeLa cell nuclear extract (substrate) and p300 (specific HAT domain region used as a positive control). Once the HAT assay has been successfully performed with the putative HAT enzymes, a variety of substrates (i.e. histones purified from S. cerevisiae and C. neoformans, synthetic chromatin made in the lab) will be obtained and incorporated into the HAT assay protocol to further study their mechanisms of action. In addition to these putative HAT enzymes, our research aims to expand on and modify the current working HAT assay protocol in order to test putative histone deacetylases (HDACs), which perform the opposite function of HATs by removing acetyl groups from histone tails.
Examination of Putative Transcriptional Regulators in Cryptococcus Neoformans
Poster - Basic
Histone acetyltransferases (HATs) are enzymes involved in the transcriptional activation of genes. p300, a known HAT enzyme, acetylates histone tails at lysine and arginine residues. When histones are acetylated, they are more prone to promote transcriptional activation. Therefore, investigation into different HATs can aid in understanding gene regulation. Within C. neoformans, a pathogenic strain of yeast, there are a couple of putative HAT enzymes that are the focus of our research, the main one being the SAS3 gene, which shows a significant upregulation in expression when C. neoformans is allowed to grow in the absence of glucose . However, although the sequences of these genes suggest they are HAT enzymes, there is no definitive proof existing about their enzymatic activity. Our research aims to first isolate these proteins that have been cloned into a bacterial vector (pet45b(+)) via a His-tag purification method. Once isolated, the proteins can be assessed for their potential HAT activity via HAT assay. Every HAT assay requires an Acetyl-CoA source, substrate, and enzyme. Our current working HAT assay protocol involves using core histones purified from HeLa cell nuclear extract (substrate) and p300 (specific HAT domain region used as a positive control). Once the HAT assay has been successfully performed with the putative HAT enzymes, a variety of substrates (i.e. histones purified from S. cerevisiae and C. neoformans, synthetic chromatin made in the lab) will be obtained and incorporated into the HAT assay protocol to further study their mechanisms of action. In addition to these putative HAT enzymes, our research aims to expand on and modify the current working HAT assay protocol in order to test putative histone deacetylases (HDACs), which perform the opposite function of HATs by removing acetyl groups from histone tails.
Comments
Undergraduate