The Dual Effect of Mimosine on DNA Replication

Abstract

The plant amino acid, mimosine, is an extremely effective inhibitor of DNA replication in mammalian cells, but the mechanism by which this inhibition is achieved is unknown. The drug has been proposed either to inhibit initiation at origins of replication or to inhibit chain elongation by lowering nucleotide pool levels. In an attempt to determine which mode of action is correct, we have analyzed its effects on SV40 DNA replication. Using a two-dimensional gel replicon mapping technique, we show that mimosine completely inhibits incorporation of [ 3H]thymidine into viral DNA, but only after ∼4 h. Qualitative analysis of replication intermediates during this interval suggests that the drug partially inhibits both initiation and elongation, and pulse–chase experiments support this contention. The drug has no effect when added directly to an SV40 in vitroreplication extract. However, extracts prepared from cells pretreated with mimosine are compromised in their ability to support replication in vitroin the presence of a full complement of nucleotides. Thus, although mimosine may alter nucleotide pool levels in vivo,it also appears to affect one or more essential replication proteins.