Figure 9

Nuclease P1 cleavage of total tRNA from yfhQ K.O. strain incubated with YfhQ and AdoMet. Autoradiography of a two-dimensional chromatogram of 5'-phosphate nucleosides on a thin layer cellulose plate. The reaction mixture containing 50 mM PIPES-Na, pH 7.0, 4 mM MgCl₂, 100 μg of total tRNA from yfhQ K.O. strain, 5 μg of the purified YfhQ protein, and 10 μM [¹⁴C-methyl]AdoMet (53 mCi/mmol). After 60 min incubation at 37°C, the tRNA was recovered, digested by nuclease P1 and the resulting nucleotides were analyzed as described [38].