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Evolutionary Robustness of an Optimal Phenotype: Re-evolution of Lysis in a Bacteriophage Deleted for Its Lysin Gene

Journal of Molecular Evolution volume 61, pages 181–191 (2005)Cite this article

Abstract

Optimality models are frequently used to create expectations about phenotypic evolution based on the fittest possible phenotype. However, they often ignore genetic details, which could confound these expectations. We experimentally analyzed the ability of organisms to evolve towards an optimum in an experimentally tractable system, lysis time in bacteriophage T7. T7 lysozyme helps lyse the host cell by degrading its cell wall at the end of infection, allowing viral escape to infect new hosts. Artificial deletion of lysozyme greatly reduced fitness and delayed lysis, but after evolution both phenotypes approached wild-type values. Phage with a lysis-deficient lysozyme evolved similarly. Several mutations were involved in adaptation, but most of the change in lysis timing and fitness increase was mediated by changes in gene 16, an internal virion protein not formerly considered to play a role in lysis. Its muralytic domain, which normally aids genome entry through the cell wall, evolved to cause phage release. Theoretical models suggest there is an optimal lysis time, and lysis more rapid or delayed than this optimum decreases fitness. Artificially constructed lines with very rapid lysis had lower fitness than wild-type T7, in accordance with the model. However, while a slow-lysing line also had lower fitness than wild-type, this low fitness resulted at least partly from genetic details that violated model assumptions.

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References

  • Abedon ST (1992) Lysis of lysis-inhibited bacteriophage T4-infected cells. J Bacteriol 174:8073–8080

    PubMed  Google Scholar 

  • Abedon ST, Herschler TD, Stopar D (2001) Bacteriophage latent-period evolution as a response to resource availability. Appl Environ Microbiol 67:233–4241

    Google Scholar 

  • Abedon ST, Hyman P, Thomas C (2003) Experimental examination of bacteriophage latent-period evolution as a response to bacterial availability. Appl Environ Microbiol 69:7499–7506

    Article  PubMed  Google Scholar 

  • Bull JJ, Badgett MR, Rokyta D, Molineux IJ (2003) Experimental evolution yields hundreds of mutations in a functional viral genome. J Mol Evol 57:241–248

    Article  PubMed  Google Scholar 

  • Bull JJ, Badgett MR, Wichman HA (2000) Big-benefit mutations in a bacteriophage inhibited with heat. Mol Biol Evol 17:942–950

    Google Scholar 

  • Chang CY, Nam K, Young R (1995) S gene expression and the timing of lysis by bacteriophage lambda. J Bacteriol 177:3283–3294

    PubMed  Google Scholar 

  • Charnov EL (1982) The theory of sex allocation. Monogr Popul Biol 18:1–355

    PubMed  Google Scholar 

  • Cheng X, Zhang X, Pflugrath JW, Studier FW (1994) The structure of bacteriophage T7 lysozyme, a zinc amidase and an inhibitor of T7 RNA polymerase. Proc Natl Acad Sci USA 91:4034–4038

    PubMed  Google Scholar 

  • Dunn JJ, Studier FW (1983) Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol 166:477–535

    Google Scholar 

  • Engel H, Kazemier B, Keck W (1991) Murein-metabolizing enzymes from Escherichia coli: sequence analysis and controlled overexpression of the slt gene, which encodes the soluble lytic transglycosylase. J Bacteriol 173:6773–6782

    PubMed  Google Scholar 

  • Freeland SJ, Knight RD, Landweber LF, Hurst LD (2000) Early fixation of an optimal genetic code. Mol Biol Evol 17:511–518

    PubMed  Google Scholar 

  • Garcia LR, Molineux IJ (1996) Transcription-independent DNA translocation of bacteriophage T7 DNA into Escherichia coli. J Bacteriol 178:6921–6929

    PubMed  Google Scholar 

  • Hutchinson CA, Sinsheimer RL (1966) The process of infection with bacteriophage phiX174. J. Mol Biol. 18:429–447

    PubMed  Google Scholar 

  • Inouye M, Arnheim N, Sternglanz R (1973) Bacteriophage T7 lysozyme is an N-acetylmuramyl-L-alanine amidase. J Biol Chem 248:7247-7252

    Google Scholar 

  • Josslin R (1970) The lysis mechanism of phage T4: mutants affecting lysis. Virology 40:719–726

    Article  PubMed  Google Scholar 

  • Kanamaru S, Ishiwata Y, Suzuki T, Rossmann MG, Arisaka F (2005) Control of bacteriophage T4 tail lysozyme activity during the infection process. J Mol Biol 346:1013–1020

    Article  PubMed  Google Scholar 

  • Kao SH, McClain WH (1980a) Baseplate protein of bacteriophage T4 with both structural and lytic functions. J Virol 34:95–103

    Google Scholar 

  • Kao SH, McClain WH (1980b) Roles of bacteriophage T4 gene 5 and gene s products in cell lysis. J Virol 34:104–107

    Google Scholar 

  • Kemp P, Gupta M, Molineux IJ (2004) Bacteriophage T7 DNA ejection into cells is initiated by an enzyme-like mechanism. Mol Microbiol 53:1251–1265

    Article  PubMed  Google Scholar 

  • Lehman N (2004) Assessing the likelihood of recurrence during RNA evolution in vitro. Artif Life 10:1–22

    Article  PubMed  Google Scholar 

  • Lewontin RC (1989) A natural selection. Nature 339:107

    Article  Google Scholar 

  • Lyakhov DL, He B, Zhang X, Studier FW, Dunn JJ, McAllister WT (1997) Mutant bacteriophage T7 RNA polymerases with altered termination properties. J Mol Biol 269:28–40

    Article  PubMed  Google Scholar 

  • McAllister WT, Wu HL (1978) Regulation of transcription of the late genes of bacteriophage T7. Proc Natl Acad Sci USA 75:804–808

    PubMed  Google Scholar 

  • Moak M, Molineux IJ (2000) Role of the Gp16 lytic transglycosylase motif in bacteriophage T7 virions at the initiation of infection. Mol Microbiol 37:345–355

    Article  PubMed  Google Scholar 

  • Moak M, Molineux IJ (2004) Peptidoglycan hydrolytic activities associated with bacteriophage virions. Mol Microbiol 51:1169–1183

    Article  PubMed  Google Scholar 

  • Moffat BA, Studier FW (1987) T7 lysozyme inhibits transcription by T7 RNA polymerase. Cell 49:221–227

    Article  PubMed  Google Scholar 

  • Molineux IJ (1999) T7 bacteriophages. In: Creighton TE (ed) Encyclopedia of molecular biology. New York: Wiley. p 2495–2507

    Google Scholar 

  • Molineux IJ (2001) No syringes please, ejection of phage T7 DNA from the virion is enzyme driven. Mol Microbiol 40:1–8

    Article  PubMed  Google Scholar 

  • Nakagawa H, Arisaka F, Ishii S (1985) Isolation and characterization of the bacteriophage T4 tail-associated lysozyme. J Virol 54:460–466

    PubMed  Google Scholar 

  • Rennell D, Bouvier SE, Hardy LW, Poteete AR (1991) Systematic mutation of bacteriophage T4 lysozyme. J Mol Biol 222:67–88

    Google Scholar 

  • Rokyta D, Badgett MR, Molineux IJ, Bull JJ (2002) Experimental genomic evolution: extensive compensation for loss of DNA ligase activity in a virus. Mol Biol Evol 19:230–238

    Google Scholar 

  • Scholl D, Kieleczawa J, Kemp P, Rush J, Richardson CC, Merril C, Adhya S, Molineux IJ (2004) Genomic analysis of bacteriophages SP6 and K1-5, an estranged subgroup of the T7 supergroup. J Mol Biol 335:1151–1171

    Google Scholar 

  • Silberstein S, Inouye M (1975) Studies on the role of bacteriophage T7 lysozyme during phage infection. J Mol Biol 96:1–11

    Article  PubMed  Google Scholar 

  • Smith JM (1983) Models of Evolution. Proc R Soc Lond B Biol Sci 219:315–325

    Google Scholar 

  • Studier FW (1969) The genetics and physiology of bacteriophage T7. Virology 39:562–574

    Article  PubMed  Google Scholar 

  • Studier FW (1973) Genetic analysis of non-essential bacteriophage T7 genes. J Mol Biol 79:227–236

    Article  PubMed  Google Scholar 

  • Takeda S, Hoshida K, Arisaka F (1998) Mapping of functional sites on the primary structure of the tail lysozyme of bacteriophage T4 by mutational analysis. Biochim Biophys Acta 1384:243–252

    PubMed  Google Scholar 

  • Trivers RL (1983) The evolution of sex. Q. Rev Biol 58:62–67

    Article  Google Scholar 

  • Villemain J, Sousa R (1998) Specificity in transcriptional regulation in the absence of specific DNA binding sites: the case of T7 lysozyme. J Mol Biol 281:793–802

    Google Scholar 

  • Wang IN, Dykhuizen DE, Slobodkin LB (1996) The evolution of phage lysis timing. Evol Ecol 10:545–558

    Google Scholar 

  • Wang IN, Smith DL, Young R (2000) Holins: the protein clocks of bacteriophage infections. Annu Rev Microbiol 54:799–825

    Article  PubMed  Google Scholar 

  • Williams GC (1966) Adaptation and natural selection. Princeton: Princeton University Press

    Google Scholar 

  • Yang XM, Richardson CC (1997) Amino acid changes in a unique sequence of bacteriophage T7 DNA polymerase alter the processivity of nucleotide polymerization. J Biol Chem 272:6599–6606

    Google Scholar 

  • Young I, Wang I, Roof WD (2000) Phages will out: strategies of host cell lysis. Trends Microbiol 8:120–128

    Article  PubMed  Google Scholar 

  • Young R (1992) Bacteriophage lysis: mechanism and regulation. Microbiol Rev 56:430–481

    PubMed  Google Scholar 

  • Zhang X, Studier FW (1995) Isolation of transcriptionally active mutants of T7 RNA polymerase that do not support phage growth. J Mol Biol 250:156–168

    Google Scholar 

  • Zhang X, Studier FW (1995) Multiple roles of T7RNA polymerase T7 lysozyme during bacteriophage T7 infection. J Mol Biol 340:707–730

    Google Scholar 

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Acknowledgments

We thank X. Zhang, F.W. Studier, and A.R. Poteete for phages and plasmids used in this work. Helpful suggestions were provided by R. Young, S. Hedtke, I. Wang, two anonymous reviewers, and the Bull lab group. This study was funded by NIH GM 57756 to J.J.B. and GM 32095 to I.J.M.

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Authors and Affiliations

  1. Section of Integrative Biology, University of Texas, Austin, Texas, 78712, USA

    Richard H. Heineman & James J. Bull

  2. Section of Molecular Genetics and Microbiology, University of Texas, Austin, Texas, 78712, USA

    Ian J. Molineux

  3. Institute for Cell and Molecular Biology, University of Texas, Austin, Texas, 78712, USA

    Ian J. Molineux & James J. Bull

Authors
  1. Richard H. Heineman
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  2. Ian J. Molineux
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  3. James J. Bull
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Correspondence to Richard H. Heineman.

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Heineman, R.H., Molineux, I.J. & Bull, J.J. Evolutionary Robustness of an Optimal Phenotype: Re-evolution of Lysis in a Bacteriophage Deleted for Its Lysin Gene. J Mol Evol 61, 181–191 (2005). https://doi.org/10.1007/s00239-004-0304-4

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  • DOI: https://doi.org/10.1007/s00239-004-0304-4

Keywords

  • Optimality
  • Experimental evolution
  • Evolutionary robustness
  • Lysis
  • T7
  • Bacteriophage
  • Genome evolution
  • Molecular evolution
  • Fitness
  • Adaptation