Class
Article
College
College of Science
Department
Chemistry and Biochemistry Department
Faculty Mentor
Joan M. Hevel
Presentation Type
Poster Presentation
Abstract
Protein arginine methylation is an essential post-translational modification that significantly affects various eukaryotic cellular processes by mediating protein-protein, protein-RNA, and protein-DNA interactions. Arginine methylation is catalyzed by a family of proteins called protein arginine methyltransferases (PRMTs). Proper regulation of PRMT activity is imperative, as dysregulation is found to contribute to cardiovascular disease and cancer. In the study described herein, we investigated how phosphorylation of PRMT1 might affect both the oligomerization and activity of the enzyme. A recent study demonstrated that the yeast analog of yPRMT1 (yPRMT1) can be phosphorylated at serine residue 9. Using in vivo studies, the authors claimed that phosphorylation promotes oligomerization and, consequently, increased yPRMT1 activity. However, the experimental techniques failed to provide sufficient evidence to support their claim that the phosphorylated residue intrinsically affects the active oligomeric state. Additionally, preliminary studies from our lab suggest that phosphorylation does not directly affect activity. Our study will establish whether phosphorylation has a direct effect on yPRMT1 oligomerization and activity, using conclusive experimental methods. We recombinantly expressed and purified wild-type (WT), a serine to glutamate (S9E) mutant that acts as a phosphomimic, and a serine to alanine mutant (S9A) yPRMT1 constructs using nickel affinity chromatography. Further, we will quantify methyltransferase activity using kinetic assays, as well as the oligomeric states of each of the constructs by combining data from Analytical Ultracentrifugation (AUC) and Native-PAGE. Collectively, our results suggest that phosphorylation of Ser9 does not directly affect the oligomerization state and activity of yPRMT1.
Location
Logan, UT
Start Date
4-13-2021 12:00 AM
Included in
Investigating the Direct Effect of Phosphorylation on the Oligomeric State and Activity of yPRMT1
Logan, UT
Protein arginine methylation is an essential post-translational modification that significantly affects various eukaryotic cellular processes by mediating protein-protein, protein-RNA, and protein-DNA interactions. Arginine methylation is catalyzed by a family of proteins called protein arginine methyltransferases (PRMTs). Proper regulation of PRMT activity is imperative, as dysregulation is found to contribute to cardiovascular disease and cancer. In the study described herein, we investigated how phosphorylation of PRMT1 might affect both the oligomerization and activity of the enzyme. A recent study demonstrated that the yeast analog of yPRMT1 (yPRMT1) can be phosphorylated at serine residue 9. Using in vivo studies, the authors claimed that phosphorylation promotes oligomerization and, consequently, increased yPRMT1 activity. However, the experimental techniques failed to provide sufficient evidence to support their claim that the phosphorylated residue intrinsically affects the active oligomeric state. Additionally, preliminary studies from our lab suggest that phosphorylation does not directly affect activity. Our study will establish whether phosphorylation has a direct effect on yPRMT1 oligomerization and activity, using conclusive experimental methods. We recombinantly expressed and purified wild-type (WT), a serine to glutamate (S9E) mutant that acts as a phosphomimic, and a serine to alanine mutant (S9A) yPRMT1 constructs using nickel affinity chromatography. Further, we will quantify methyltransferase activity using kinetic assays, as well as the oligomeric states of each of the constructs by combining data from Analytical Ultracentrifugation (AUC) and Native-PAGE. Collectively, our results suggest that phosphorylation of Ser9 does not directly affect the oligomerization state and activity of yPRMT1.