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The Population and Evolutionary Dynamics of Phage and Bacteria with CRISPR–Mediated Immunity


Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), together with associated genes (cas), form the CRISPR–cas adaptive immune system, which can provide resistance to viruses and plasmids in bacteria and archaea. Here, we use mathematical models, population dynamic experiments, and DNA sequence analyses to investigate the host–phage interactions in a model CRISPR–cas system, Streptococcus thermophilus DGCC7710 and its virulent phage 2972. At the molecular level, the bacteriophage-immune mutant bacteria (BIMs) and CRISPR–escape mutant phage (CEMs) obtained in this study are consistent with those anticipated from an iterative model of this adaptive immune system: resistance by the addition of novel spacers and phage evasion of resistance by mutation in matching sequences or flanking motifs. While CRISPR BIMs were readily isolated and CEMs generated at high rates (frequencies in excess of 10−6), our population studies indicate that there is more to the dynamics of phage–host interactions and the establishment of a BIM–CEM arms race than predicted from existing assumptions about phage infection and CRISPR–cas immunity. Among the unanticipated observations are: (i) the invasion of phage into populations of BIMs resistant by the acquisition of one (but not two) spacers, (ii) the survival of sensitive bacteria despite the presence of high densities of phage, and (iii) the maintenance of phage-limited communities due to the failure of even two-spacer BIMs to become established in populations with wild-type bacteria and phage. We attribute (i) to incomplete resistance of single-spacer BIMs. Based on the results of additional modeling and experiments, we postulate that (ii) and (iii) can be attributed to the phage infection-associated production of enzymes or other compounds that induce phenotypic phage resistance in sensitive bacteria and kill resistant BIMs. We present evidence in support of these hypotheses and discuss the implications of these results for the ecology and (co)evolution of bacteria and phage.


Vyšlo v časopise: The Population and Evolutionary Dynamics of Phage and Bacteria with CRISPR–Mediated Immunity. PLoS Genet 9(3): e32767. doi:10.1371/journal.pgen.1003312
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.pgen.1003312

Souhrn

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), together with associated genes (cas), form the CRISPR–cas adaptive immune system, which can provide resistance to viruses and plasmids in bacteria and archaea. Here, we use mathematical models, population dynamic experiments, and DNA sequence analyses to investigate the host–phage interactions in a model CRISPR–cas system, Streptococcus thermophilus DGCC7710 and its virulent phage 2972. At the molecular level, the bacteriophage-immune mutant bacteria (BIMs) and CRISPR–escape mutant phage (CEMs) obtained in this study are consistent with those anticipated from an iterative model of this adaptive immune system: resistance by the addition of novel spacers and phage evasion of resistance by mutation in matching sequences or flanking motifs. While CRISPR BIMs were readily isolated and CEMs generated at high rates (frequencies in excess of 10−6), our population studies indicate that there is more to the dynamics of phage–host interactions and the establishment of a BIM–CEM arms race than predicted from existing assumptions about phage infection and CRISPR–cas immunity. Among the unanticipated observations are: (i) the invasion of phage into populations of BIMs resistant by the acquisition of one (but not two) spacers, (ii) the survival of sensitive bacteria despite the presence of high densities of phage, and (iii) the maintenance of phage-limited communities due to the failure of even two-spacer BIMs to become established in populations with wild-type bacteria and phage. We attribute (i) to incomplete resistance of single-spacer BIMs. Based on the results of additional modeling and experiments, we postulate that (ii) and (iii) can be attributed to the phage infection-associated production of enzymes or other compounds that induce phenotypic phage resistance in sensitive bacteria and kill resistant BIMs. We present evidence in support of these hypotheses and discuss the implications of these results for the ecology and (co)evolution of bacteria and phage.


Zdroje

1. BarrangouR, FremauxC, DeveauH, RichardsM, BoyavalP, et al. (2007) CRISPR provides acquired resistance against viruses in prokaryotes. Science 315: 1709–1712.

2. MarraffiniLA, SontheimerEJ (2008) CRISPR interference limits horizontal gene transfer in staphylococci by targeting DNA. Science 322: 1843–1845.

3. DeveauH, GarneauJE, MoineauS (2010) CRISPR/Cas system and its role in phage-bacteria interactions. Annu Rev Microbiol 64: 475–493.

4. HaleCR, ZhaoP, OlsonS, DuffMO, GraveleyBR, et al. (2009) RNA-guided RNA cleavage by a CRISPR RNA-Cas protein complex. Cell 139: 945–956.

5. BrounsSJ, JoreMM, LundgrenM, WestraER, SlijkhuisRJ, et al. (2008) Small CRISPR RNAs guide antiviral defense in prokaryotes. Science 321: 960–964.

6. DeltchevaE, ChylinskiK, SharmaCM, GonzalesK, ChaoY, et al. (2011) CRISPR RNA maturation by trans-encoded small RNA and host factor RNase III. Nature 471: 602–607.

7. DeveauH, BarrangouR, GarneauJE, LabonteJ, FremauxC, et al. (2008) Phage response to CRISPR–encoded resistance in Streptococcus thermophilus. J Bacteriol 190: 1390–1400.

8. GarneauJE, DupuisME, VillionM, RomeroDA, BarrangouR, et al. (2010) The CRISPR/Cas bacterial immune system cleaves bacteriophage and plasmid DNA. Nature 468: 67–71.

9. SapranauskasR, GasiunasG, FremauxC, BarrangouR, HorvathP, et al. (2011) The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res 39: 9275–9282.

10. AnderssonAF, BanfieldJF (2008) Virus population dynamics and acquired virus resistance in natural microbial communities. Science 320: 1047–1050.

11. TysonGW, BanfieldJF (2008) Rapidly evolving CRISPRs implicated in acquired resistance of microorganisms to viruses. Environ Microbiol 10: 200–207.

12. SnyderJC, BatesonMM, LavinM, YoungMJ (2009) Use of cellular CRISPR (clusters of regularly interspaced short palindromic repeats) spacer-based microarrays for detection of viruses in environmental samples. Appl Environ Microbiol 76: 7251–7258.

13. BrudeyK, DriscollJR, RigoutsL, ProdingerWM, GoriA, et al. (2006) Mycobacterium tuberculosis complex genetic diversity: mining the fourth international spoligotyping database (SpolDB4) for classification, population genetics and epidemiology. BMC Microbiol 6: 23.

14. CuiY, LiY, GorgeO, PlatonovME, YanY, et al. (2008) Insight into microevolution of Yersinia pestis by clustered regularly interspaced short palindromic repeats. PLoS ONE 3: e2652 doi:10.1371/journal.pone.0002652.

15. PourcelC, SalvignolG, VergnaudG (2005) CRISPR elements in Yersinia pestis acquire new repeats by preferential uptake of bacteriophage DNA, and provide additional tools for evolutionary studies. Microbiology 151: 653–663.

16. VergnaudG, LiY, GorgeO, CuiY, SongY, et al. (2007) Analysis of the three Yersinia pestis CRISPR loci provides new tools for phylogenetic studies and possibly for the investigation of ancient DNA. Adv Exp Med Biol 603: 327–338.

17. LiuF, BarrangouR, Gerner-SmidtP, RibotEM, KnabelSJ, et al. (2011) Novel virulence gene and clustered regularly interspaced short palindromic repeat (CRISPR) multilocus sequence typing scheme for subtyping of the major serovars of Salmonella enterica subsp. enterica. Appl Environ Microbiol 77: 1946–1956.

18. BarrangouR, HorvathP (2012) CRISPR: New Horizons in Phage Resistance and Strain Identification. Annu Rev Food Sci Technol 3: 143–162.

19. PalmerKL, GilmoreMS (2010) Multidrug-resistant enterococci lack CRISPR–cas. MBio 1: e00227–10 doi:10.1128/mBio.00227-10.

20. BolotinA, QuinquisB, SorokinA, EhrlichSD (2005) Clustered regularly interspaced short palindrome repeats (CRISPRs) have spacers of extrachromosomal origin. Microbiology 151: 2551–2561.

21. MojicaFJ, Diez-VillasenorC, Garcia-MartinezJ, SoriaE (2005) Intervening sequences of regularly spaced prokaryotic repeats derive from foreign genetic elements. J Mol Evol 60: 174–182.

22. LintnerNG, KerouM, BrumfieldSK, GrahamS, LiuH, et al. Structural and functional characterization of an archaeal clustered regularly interspaced short palindromic repeat (CRISPR)-associated complex for antiviral defense (CASCADE). J Biol Chem 286: 21643–21656.

23. RhoM, WuYW, TangH, DoakTG, YeY (2012) Diverse CRISPRs evolving in human microbiomes. PLoS Genet 8: e1002441 doi:10.1371/journal.pgen.1002441.

24. LevinBR (2010) Nasty viruses, costly plasmids, population dynamics, and the conditions for establishing and maintaining CRISPR–mediated adaptive immunity in bacteria. PLoS Genet 6: e1001171 doi:10.1371/journal.pgen.1001171.

25. ChildsL, HeildN, YoungMJ, WhitakerRJ, WeitzJS (2012) Mult-scale model of CRISRP-induced coevolutionary dynamics: Diversification at the inteface of Lamarck and Darwin. Evolution doi:10.1111/j.1558-5646.2012.01595.x.

26. WeinbergerA, SunC, PlucińskiM, DeneV, ThomasB, et al. (2012) Persisting Viral Sequences Shape Microbial CRISPR–based Immunity. PLoS Comput Biol 8: e1002475 doi:10.1371/journal.pcbi.1002475.

27. ValePF, LittleTJ (2010) CRISPR–mediated phage resistance and the ghost of coevolution past. Proc Biol Sci doi:10.1098/rspb.2010.0055.

28. Adams MH (1959) Bacteriophages. New York: Wiley-Interscience.

29. MonodJ (1949) The growth of bacterial cultures. Annual Review of Microbiology 3: 371–394.

30. StewartFM, LevinBR (1973) Resource partitioning and the outcome of interspecific competition: a model and some general considerations. American Naturalist 107: 171–198.

31. LevinBR, StewartFM, ChaoL (1977) Resource-Limited Growth, Competition, and Predation - a Model and Experimental Studies with Bacteria and Bacteriophage. American Naturalist 111: 3–24.

32. DuplessisM, RussellWM, RomeroDA, MoineauS (2005) Global gene expression analysis of two Streptococcus thermophilus bacteriophages using DNA microarray. Virology 340: 192–208.

33. DeveauH, Van CalsterenMR, MoineauS (2002) Effect of exopolysaccharides on phage-host interactions in Lactococcus lactis. Appl Environ Microbiol 68: 4364–4369.

34. LevesqueC, DuplessisM, LabonteJ, LabrieS, FremauxC, et al. (2005) Genomic organization and molecular analysis of virulent bacteriophage 2972 infecting an exopolysaccharide-producing Streptococcus thermophilus strain. Appl Environ Microbiol 71: 4057–4068.

35. BarrangouR, FremauxC, DeveauH, RichardsM, BoyavalP, et al. (2007) CRISPR provides acquired resistance against viruses in prokaryotes. Science 315: 1709–1712.

36. HorvathP, RomeroDA, Coute-MonvoisinAC, RichardsM, DeveauH, et al. (2008) Diversity, activity, and evolution of CRISPR loci in Streptococcus thermophilus.. J Bacteriol 190: 1401–1412.

37. BiggerJW (1944) Treatment of staphylococcal infections with penicillin - By intermittent sterilisation. Lancet 2: 497–500.

38. LiK, BarksdaleL, GarmiseL (1961) Phenotypic alterations associated with the bacteriophage carrier state of Shigella dysenteriae. J Gen Microbiol 24: 355–367.

39. BarksdaleL, ArdenSB (1974) Persisting bacteriophage infections, lysogeny, and phage conversions. Annu Rev Microbiol 28: 265–299.

40. StewartFM, LevinBR (1977) The population biology of bacterial plasmids: a priori conditions for the existence of conjugationally transmitted factors. Genetics 87: 209–228.

41. SunCL, BarrangouR, ThomasBC, HorvathP, FremauxC, et al. (2012) Phage mutations in response to CRISPR diversification in a bacterial population. Environ Microbiol 15 (2)

463–70 doi:10.1111/j.1462-2920.2012.02879.x.

42. LenskiRE, LevinBR (1985) Constraints on the Coevolution of Bacteria and Virulent Phage - a Model, Some Experiments, and Predictions for Natural Communities. American Naturalist 125: 585–602.

43. MeyerJR, DobiasDT, WeitzJS, BarrickJE, QuickRT, et al. (2012) Repeatability and contingency in the evolution of a key innovation in phage lambda. Science 335: 428–432.

44. TremblayDM, MoineauS (1999) Complete genomic sequence of the lytic bacteriophage DT1 of Streptococcus thermophilus. Virology 255: 63–76.

Štítky
Genetika Reprodukčná medicína

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PLOS Genetics


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