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Mechanistic Investigations Of Pseudouridine Synthases : A Surprising Glycal Intermediate Lies On The Reaction Pathway., Govardhan Reddy Veerareddygari
Mechanistic Investigations Of Pseudouridine Synthases : A Surprising Glycal Intermediate Lies On The Reaction Pathway., Govardhan Reddy Veerareddygari
Electronic Theses and Dissertations
Pseudouridine synthases (ΨSs) catalyze the isomerization of uridine (U) in RNA to pseudouridine (Ψ), which is the most common post-transcriptional modification in RNAs and is ubiquitous within all three domains of life. ΨSs are classified into six different families based on sequence alignments and have a universally conserved aspartic acid residue (‘conserved Asp’ for simplicity) that is absolutely essential for activity. RNA containing 5-fluorouridine ([F5U]RNA) has been used as a mechanistic probe. Upon incubation with [F5U]RNA, TruA and RluA get irreversibly inhibited and appear in a protein-RNA adduct band on denaturing PAGE gels, which is consistent with the ‘Michael mechanism’ …