Human immunodeficiency virus reverse transcriptase substitutes for DNA polymerase I in Escherichia coli

Proc Natl Acad Sci U S A. 1995 Jan 31;92(3):684-8. doi: 10.1073/pnas.92.3.684.


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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • DNA Polymerase I / metabolism*
  • DNA Replication / drug effects
  • Drug Evaluation, Preclinical
  • Escherichia coli / drug effects
  • Escherichia coli / enzymology*
  • Escherichia coli / genetics
  • Genetic Complementation Test
  • HIV / enzymology*
  • Humans
  • Mutation / physiology
  • Plasmids
  • RNA-Directed DNA Polymerase / metabolism*
  • Temperature
  • Zidovudine / pharmacology


  • Zidovudine
  • RNA-Directed DNA Polymerase
  • DNA Polymerase I