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. 2000 Dec;156(4):1471–1481. doi: 10.1093/genetics/156.4.1471

Compensatory evolution in rifampin-resistant Escherichia coli.

M G Reynolds 1
PMCID: PMC1461348  PMID: 11102350

Abstract

This study examines the intrinsic fitness burden associated with RNA polymerase (rpoB) mutations conferring rifampin resistance in Escherichia coli K12 (MG1655) and explores the nature of adaptation to the costs of resistance. Among 28 independent Rif(r) mutants, the per-generation fitness burden (in the absence of rifampin) ranged from 0 to 28%, with a median of 6.4%. We detected no relationship between the magnitude of the cost and the level of resistance. Adaptation to the costs of rif resistance was studied by following serial transfer cultures for several Rif(r) mutants both in the presence of rifampin and in the absence. For cultures evolved in the absence of rifampin, single clones isolated after 200 generations were more fit than their ancestor; we saw no association between increased fitness and changes in the level of rifampin resistance; and in all cases, increased fitness was due to compensatory mutations, rather than to reversion to drug sensitivity. However, in the parallel evolution experiments in the presence of rifampin, overall levels of resistance increased as did relative fitness-for all strains save one that had an initially high level of resistance. Among the evolved clones tested, five (of seven) demonstrated increased transcription efficiency (assessed using a semiquantitative RT-PCR protocol). The implications of these results for our understanding of adaptive molecular evolution and the increasing clinical problem of antibiotic resistance are discussed.

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Selected References

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  1. Aubry-Damon H., Soussy C. J., Courvalin P. Characterization of mutations in the rpoB gene that confer rifampin resistance in Staphylococcus aureus. Antimicrob Agents Chemother. 1998 Oct;42(10):2590–2594. doi: 10.1128/aac.42.10.2590. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Billington O. J., McHugh T. D., Gillespie S. H. Physiological cost of rifampin resistance induced in vitro in Mycobacterium tuberculosis. Antimicrob Agents Chemother. 1999 Aug;43(8):1866–1869. doi: 10.1128/aac.43.8.1866. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Björkman J., Hughes D., Andersson D. I. Virulence of antibiotic-resistant Salmonella typhimurium. Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3949–3953. doi: 10.1073/pnas.95.7.3949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Björkman J., Nagaev I., Berg O. G., Hughes D., Andersson D. I. Effects of environment on compensatory mutations to ameliorate costs of antibiotic resistance. Science. 2000 Feb 25;287(5457):1479–1482. doi: 10.1126/science.287.5457.1479. [DOI] [PubMed] [Google Scholar]
  5. Björkman J., Samuelsson P., Andersson D. I., Hughes D. Novel ribosomal mutations affecting translational accuracy, antibiotic resistance and virulence of Salmonella typhimurium. Mol Microbiol. 1999 Jan;31(1):53–58. doi: 10.1046/j.1365-2958.1999.01142.x. [DOI] [PubMed] [Google Scholar]
  6. Borman A. M., Paulous S., Clavel F. Resistance of human immunodeficiency virus type 1 to protease inhibitors: selection of resistance mutations in the presence and absence of the drug. J Gen Virol. 1996 Mar;77(Pt 3):419–426. doi: 10.1099/0022-1317-77-3-419. [DOI] [PubMed] [Google Scholar]
  7. Filutowicz M. Requirement of DNA gyrase for the initiation of chromosome replication in Escherichia coli K-12. Mol Gen Genet. 1980 Jan;177(2):301–309. doi: 10.1007/BF00267443. [DOI] [PubMed] [Google Scholar]
  8. Foley K. P., Leonard M. W., Engel J. D. Quantitation of RNA using the polymerase chain reaction. Trends Genet. 1993 Nov;9(11):380–385. doi: 10.1016/0168-9525(93)90137-7. [DOI] [PubMed] [Google Scholar]
  9. Gong M., Cowan K. H., Gudas J., Moscow J. A. Isolation and characterization of genomic sequences involved in the regulation of the human reduced folate carrier gene (RFC1). Gene. 1999 Jun 11;233(1-2):21–31. doi: 10.1016/s0378-1119(99)00166-3. [DOI] [PubMed] [Google Scholar]
  10. Guyer M. S., Reed R. R., Steitz J. A., Low K. B. Identification of a sex-factor-affinity site in E. coli as gamma delta. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):135–140. doi: 10.1101/sqb.1981.045.01.022. [DOI] [PubMed] [Google Scholar]
  11. Herrera G., Aleixandra V., Urios A., Blanco M. Quinolone action in Escherichia coli cells carrying gyrA and gyrB mutations. FEMS Microbiol Lett. 1993 Jan 15;106(2):187–191. doi: 10.1016/0378-1097(93)90079-h. [DOI] [PubMed] [Google Scholar]
  12. Houze T. A., Larsson L., Larsson P. A., Hansson G., Asea A., Gustavsson B. Rapid detection of thymidylate synthase gene expression levels by semi-quantitative competitive reverse transcriptase polymerase chain reaction followed by quantitative digital image analysis. Tumour Biol. 1996;17(5):306–319. doi: 10.1159/000217993. [DOI] [PubMed] [Google Scholar]
  13. Moore F. B., Rozen D. E., Lenski R. E. Pervasive compensatory adaptation in Escherichia coli. Proc Biol Sci. 2000 Mar 7;267(1442):515–522. doi: 10.1098/rspb.2000.1030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Musser J. M. Antimicrobial agent resistance in mycobacteria: molecular genetic insights. Clin Microbiol Rev. 1995 Oct;8(4):496–514. doi: 10.1128/cmr.8.4.496. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Padayachee T., Klugman K. P. Molecular basis of rifampin resistance in Streptococcus pneumoniae. Antimicrob Agents Chemother. 1999 Oct;43(10):2361–2365. doi: 10.1128/aac.43.10.2361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Pozzi G., Meloni M., Iona E., Orrù G., Thoresen O. F., Ricci M. L., Oggioni M. R., Fattorini L., Orefici G. rpoB mutations in multidrug-resistant strains of Mycobacterium tuberculosis isolated in Italy. J Clin Microbiol. 1999 Apr;37(4):1197–1199. doi: 10.1128/jcm.37.4.1197-1199.1999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Reynolds M. G., Roos D. S. A biochemical and genetic model for parasite resistance to antifolates. Toxoplasma gondii provides insights into pyrimethamine and cycloguanil resistance in Plasmodium falciparum. J Biol Chem. 1998 Feb 6;273(6):3461–3469. doi: 10.1074/jbc.273.6.3461. [DOI] [PubMed] [Google Scholar]
  18. Schrag S. J., Perrot V., Levin B. R. Adaptation to the fitness costs of antibiotic resistance in Escherichia coli. Proc Biol Sci. 1997 Sep 22;264(1386):1287–1291. doi: 10.1098/rspb.1997.0178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schrag S. J., Perrot V. Reducing antibiotic resistance. Nature. 1996 May 9;381(6578):120–121. doi: 10.1038/381120b0. [DOI] [PubMed] [Google Scholar]
  20. Weisblum B., Davies J. Antibiotic inhibitors of the bacterial ribosome. Bacteriol Rev. 1968 Dec;32(4 Pt 2):493–528. [PMC free article] [PubMed] [Google Scholar]
  21. Yanofsky C., Horn V. Rifampin resistance mutations that alter the efficiency of transcription termination at the tryptophan operon attenuator. J Bacteriol. 1981 Mar;145(3):1334–1341. doi: 10.1128/jb.145.3.1334-1341.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]

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