MBE Advance Access originally published online on June 3, 2009
Molecular Biology and Evolution 2009 26(9):2041-2046; doi:10.1093/molbev/msp111
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Research Articles |
Viral Resistance Evolution Fully Escapes a Rationally Designed Lethal Inhibitor
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* Section of Integrative Biology, The University of Texas at Austin
Section of Molecular Genetics and Microbiology, The University of Texas at Austin
Institute for Cellular and Molecular Biology, The University of Texas at Austin
Center for Computational Biology and Bioinformatics, The University of Texas at Austin
E-mail: tkeller{at}mail.utexas.edu.
Accepted for publication May 22, 2009.
Viruses are notoriously capable of evolving resistance to drugs. However, if the endpoint of resistance evolution is only partial escape, a feasible strategy should be to stack drugs, so the combined effect of partial inhibition by several drugs results in net inhibition. Assessing the feasibility of this approach requires quantitative data on viral fitness before and after evolution of resistance to a drug, as done here with bacteriophage T7. An inhibitory gene expressed from a phage promoter aborts wild-type T7 infections. The effect is so severe that the phage population declines when exposed to the inhibitor but expands a billion-fold per hour in its absence. In prior work, T7 evolved modest resistance to this inhibitor, an expected result. Given the nature of the inhibitor, that it used the phage's own promoter to target the phage's destruction, we anticipated that resistance evolution would be limited as the phage may need to evolve a new regulatory system, with simultaneous changes in its RNA polymerase (RNAP) and many of its promoters to fully escape inhibition. We show here that further adaptation of the partially resistant phage led to complete resistance. Resistance evolution was due to three mutations in the RNAP gene and two other genes; unexpectedly, no changes were observed in promoters. Consideration of other mechanisms of T7 inhibition leaves hope that permanent inhibition of viral growth with drugs can in principle be achieved.
Key Words: drug resistance • experimental evolution • T7 bacteriophage • microbe • viral adaptation
Manolo Gouy, Associate Editor