Human immunodeficiency virus reverse transcriptase substitutes for DNA polymerase I in Escherichia coli.
We present evidence that human immunodeficiency virus (HIV) reverse transcriptase (RT) can substitute for DNA polymerase I in bacteria. Expression of HIV RT enables an Escherichia coli mutant, polA12 recA718, containing a temperature-sensitive mutation in DNA polymerase I, to grow at a nonpermissive temperature. The plasmid pBR322 contains a DNA polymerase I-dependent origin of replication. Expression of HIV RT enables the same E. coli mutant to maintain this plasmid at a nonpermissive temperature. Furthermore, expression of HIV RT in this mutant renders it sensitive to 3'-azido-3'-deoxythymidine, a commonly used anti-AIDS drug that targets HIV RT. These combined findings on the genetic complementation of DNA polymerase I by HIV RT provide a bacterial assay to screen for drugs directed against HIV RT. Genetic complementation provides a method for positive selection of large numbers of functional HIV RT mutants for studies on structure-function relationships.
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Joseph Gottstein Memorial Cancer Research Laboratory, Department of Pathology, University of Washington, Seattle 98195.
Loeb
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D004256:DNA Polymerase I / Q000378:metabolism*; D004261:DNA Replication / Q000187:drug effects; D004353:Drug Evaluation, Preclinical; D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D005816:Genetic Complementation Test; D006678:HIV / Q000201:enzymology*; D006801:Humans; D009154:Mutation / Q000502:physiology; D010957:Plasmids; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism*; D013696:Temperature; D015215:Zidovudine / Q000494:pharmacology
D015215:Zidovudine; D012194:RNA-Directed DNA Polymerase; D004256:DNA Polymerase I