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In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme
Published on: October 20, 2010
Enzymatic mechanism of tRNA (m5U54)methyltransferase
1Department of Pharmaceutical Chemistry, University of California, San Francisco 94143-0448, USA.
Abstract:
tRNA (m5U54)methyltransferase (RUMT) catalyzes the methylation of uridine 54 of transfer RNA by S-adenosyl-L-methionine. In this report, we present the enzymatic mechanism of RUMT, including the stereochemical course of the methylation reaction, and discuss RUMT-tRNA recognition. As part of its enzymatic mechanism, we postulate that RUMT catalyzes the disruption of RNA-RNA contacts. We also show that many nucleotide substitutions can be made in the T-loop of tRNA without affecting RUMT binding, indicating that the recognition of the T-loop by RUMT is not stringent.
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