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| Abstract |
The methylation of the sixth position of adenine (m6A) is a vital epitranscriptomic alteration that has been found to play a vital role in early cell differentiation and embryonic development. The removal of the m6A modification is facilitated by “eraser” protein FTO. The protein FTO (Fat-mass and obesity-related transcript) is a demethylase enzyme whose activity has been recently linked to cancer progression and development. However, little is known about the biophysical nature of its binding to its RNA targets. A TLC-based Mobility Shift Assay with γ32P labeled mRNA which was incubated with FTO resulted in loss of the m6A modification. Additionally, FRET experiments were used to determine the effect of FTO’s cofactors (α-ketoglutarate and Fe (II)) on its binding thermodynamics. FRET experiments also indicated the potential of a phage-display isolated peptide to serve as an inhibitor to FTO’s demethylase activity. Recently, more work has been undertaken to investigate the binding of this peptide to a modified RNA target using Circular Dichroism spectroscopy.
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| Contributor(s) |
Faculty Mentor
Sanjaya Abeysirigunawardena |
| Modified Abstract |
The methylation of the sixth position of adenine (m6A) is a vital epitranscriptomic alteration that has been found to play a vital role in cell differentiation/development. The removal of the m6A modification is facilitated by the demethylase “eraser” protein FTO, an enzyme whose activity has been recently linked to cancer progression. However, little is known about the biophysical nature of its binding to its RNA targets. A TLC-based Assay with γ32P labeled mRNA which was incubated with FTO resulted in loss of the m6A modification. Additionally, FRET experiments were used to determine the effect of FTO’s cofactors (α-ketoglutarate and Fe (II)) on its binding thermodynamics. FRET experiments also indicated the potential of a phage-display isolated peptide to serve as an inhibitor to FTO’s demethylase activity. |