Graduate Thesis Or Dissertation
 

Studies on DNA precursor metabolism in vaccinia virus-infected mamalian cells

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  • Vaccinia virus-infected animal cells have been used to study the interactions between the replication of deoxyribonucleic acid (DNA) and the biosynthesis of its nucleotide precursors. Some antimetabolites that inhibit DNA replication have as their targets enzymes of nucleotide biosynthesis. Furthermore, the disruption of nucleotide metabolism can alter the fidelity of DNA replication. The isolation of viral mutants resistant to the nucleoside analogue arabinosyl cytosine has shown that resistance to certain drugs is associated with altered fidelity of replication of DNA. Furthermore, single mutations can cause altered sensitivity to several compounds. On the other hand, selection of apparent revertants indicates that several sites can be involved in drug resistance and replication fidelity. Vaccinia virus has previously been shown to code for enzymes of DNA precursor metabolism. However, attempts to isolate mutants resistant to drugs that target thymidylate synthase and dihydrofolate reductase were not successful. In fact, stimulation of host cell DNA synthesis makes vaccinia virus extremely sensitive to the effects of the folate analogue methotrexate. Taken together these observations suggest that despite its cytoplasmic site of DNA replication and its large coding capacity, vaccinia virus depends, at least in part, on host cell enzymes to supply deoxynucleoside triphosphates for DNA replication.
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