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Investigating the Role of Arginine Methylation on Serine Phosphorylation in Histone H3
Author(s) -
Mendiola Aron Judd Perez,
Fuentes Daniel,
Mendoza Mariel,
ZuritaLopez Cecilia
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.lb64
Subject(s) - histone methyltransferase , serine , phosphorylation , methylation , histone h3 , histone methylation , biochemistry , histone h2a , histone , chemistry , biology , microbiology and biotechnology , dna methylation , gene expression , gene
We set out to investigate the effects of two PTMs, methylation and phosphorylation, with the goal of understanding the role of arginine methylation on neighboring serine phosphorylation in histone H3. Work by other labs has confirmed that arginine methylation and serine phosphorylation are involved in crosstalk within an AKT motif (RXRXXS/T) where the arginine residue is two residues away from the serine residue. Herein, we use the N‐terminal tail of histone H3 and focus on its arginine‐lysine‐serine (RKS) motif to determine whether a similar interaction is taking place. Serine‐10 hyper phosphorylation of histone H3 is a known biomarker for tumorigenesis, signaling the overexpression of proto‐oncogenes that lead to tumor formation, while arginine‐8 methylation of histone H3 is implicated in gene silencing. Preliminary results have determined that serine‐10 phosphorylation downregulates arginine‐8 methylation in histone H3 peptides. To further analyze the interaction, sequential in vitro methylation and phosphorylation reactions will be carried out using the respective protein arginine methyltransferase (PRMT) and kinase. Of the family of nine known human methyltransferases, emphasis is placed on PRMT1 and PRMT5, which are known to physiologically dimethylate histone H3. While there are several kinases that deposit the phosphate group on serine‐10, we will use mitogen and stress‐activated kinase 1 (MSK1) because it physiologically phosphorylates the serine‐10 residue and specifically targets RKS motif. Methodology for analyzing the methylation and phosphorylation interactions will be done through fluorography, western blot analysis, and thin layer chromatography analysis. Investigating the RKS motif will provide greater insights into protein function and can be a springboard for investigating these modifications in other proteins. Support or Funding Information NIH Grant #SC2GM118202