Phosphorylation effects on cancer protein's DNA binding

Abstract

This study is focused on understanding the phosphorylation sites within INI-1 DNA binding domains to understand the INI-1 protein's connections to atypical teraroid/rhadboid tumor (AT/RT), an aggressive pediatric neural cancer. A truncation (1-186 amino acids) of the INI-1 plasmid was discovered to bind DNA with the same functionality as the full-length plasmid. Other studies directed interest within this same truncation toward a phosphorylation site, serine-129, to consider as a specific active site in the mysterious functions of INI-1. This identified truncation, as well as the full-length protein, was manipulated by site directed mutagenesis. The two mutations, serine to alanine and serine to aspartic acid, serve as phosphorylated and non-phosphorylated mimics, and provide quantifiable data regarding its DNA binding activity through EMSA gels.