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5-Methyl-4-thiouridine is an unmodified nucleoside that is a member of the family of ribonucleosides. It has been shown to be a potent inhibitor of RNA polymerase and catalytic activity. 5-Methyl-4-thiouridine is converted to 5-methyluracil by the enzyme uracil phosphoribosyltransferase, which then undergoes spontaneous hydrolysis to form thymine. This compound inhibits polymerase activity in vitro and in vivo, with the IC50 value being 1.5 μM for DNA polymerase I from calf thymus and 0.1 μM for E. coli RNA polymerase.