The plant amino acid mimosine (β-N(3-hydroxy-4-pyridone)-α-amino propionic acid) is toxic for animals and reversibly inhibits the replication of animal cells in vitro. We have found that mimosine inhibits the DNA synthesis of a variety of DNA viruses, including adenovirus, parvovirus, and papovavirus and the replication of vaccinia and herpes simplex virus 1 in cells in culture. However, mimosine did not inhibit the replication of SV40 DNA in a cell-free system. Because mimosine inhibition of viral DNA synthesis was reversible by iron in the medium, we hypothesized that mimosine was inhibiting ribonucleotide reductase through its capacity to chelate the iron required in the R2 subunit of this enzyme. In support of this hypothesis, we found that mimosine-treated cells had a reduction in the pools of dGTP and dATP and that mimosine inhibited ribonucleotide reductase in vitro in an iron-dependent manner.