Abstract
Deformed Epidermal Autoregulatory Factor 1 (DEAF‐1) is a transcriptional regulator that is important for embryonic development in mice, and is linked to depression and cancer in humans. The localization of DEAF‐1 and its interaction with other proteins have been shown to affect DEAF‐1 functions. To identify post‐translational modifications of DEAF‐1 that may affect function, purified proteins from HEK293T cells were subjected to liquid chromatography‐mass spectrometry. Three phosphorylated amino acids and a methylated arginine within the DNA binding domain were identified. In vitro mutagenesis and functional assays were performed to determine if these sites could influence known activities of DEAF‐1, including: DNA binding, transcriptional regulation, localization, apoptosis, and protein‐protein interactions. In vitro assays were used to confirm cyclin dependent kinase 5 (CDK5) as the modifying enzyme of at least two of the phosphorylation sites and protein arginine methyltransferase 6 and 8 (PRMT6 and PRMT8) were found to methylate DEAF‐1. How these protein modifications of DEAF‐1 regulate its functions are being explored.
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