One Small Step for a Yeast - Microevolution within Macrophages Renders Hypervirulent Due to a Single Point Mutation

English version České info

Evolution is not limited to making new species emerge and others perish over the millennia. It is also a central force in shorter-term interactions between microbes and hosts. A good example can be found in fungi, which are an underestimated cause of human diseases. Some fungi exist as commensals, and have adapted well to life on human epithelia. But as facultative pathogens, they face a different, hostile environment. We tested the ability of C. glabrata, a pathogen closely related to baker's yeast, to adapt to macrophages. We found that by adaptation, it changed its growth type completely. This allowed the fungus to escape the phagocytes, and increased its virulence in a mouse model. Sequencing the complete genome revealed surprisingly few mutations. Further analyses allowed us to detect the single mutation responsible for the phenotype, and to recreate it in the parental strain. Our work shows that fungi can adapt to immune cells, and that this adaptation can lead to an increased virulence. Since commensals are continuously exposed to host cells, we suggest that this ability could lead to unexpected phenotype changes, including an increase in virulence potential.

Vyšlo v časopise: One Small Step for a Yeast - Microevolution within Macrophages Renders Hypervirulent Due to a Single Point Mutation. PLoS Pathog 10(10): e32767. doi:10.1371/journal.ppat.1004478
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004478

Souhrn

Zdroje

1. PerlrothJ, ChoiB, SpellbergB (2007) Nosocomial fungal infections: epidemiology, diagnosis, and treatment. Med Mycol 45: 321–346.

2. BorstA, RaimerMT, WarnockDW, MorrisonCJ, Arthington-SkaggsBA (2005) Rapid acquisition of stable azole resistance by Candida glabrata isolates obtained before the clinical introduction of fluconazole. Antimicrob Agents Chemother 49: 783–787.

3. WarnockDW, BurkeJ, CopeNJ, JohnsonEM, von FraunhoferNA, et al. (1988) Fluconazole resistance in Candida glabrata. Lancet 2: 1310.

4. SanglardD, IscherF, CalabreseD, MajcherczykPA, BilleJ (1999) The ATP binding cassette transporter gene CgCDR1 from Candida glabrata is involved in the resistance of clinical isolates to azole antifungal agents. Antimicrob Agents Chemother 43: 2753–2765.

5. RoetzerA, GabaldonT, SchullerC (2011) From Saccharomyces cerevisiae to Candida glabrata in a few easy steps: important adaptations for an opportunistic pathogen. FEMS Microbiol Lett 314: 1–9.

6. ShinJH, ChaeMJ, SongJW, JungSI, ChoD, et al. (2007) Changes in karyotype and azole susceptibility of sequential bloodstream isolates from patients with Candida glabrata candidemia. J Clin Microbiol 45: 2385–2391.

7. BaderO, SchwarzA, KraneveldEA, TangwattanchuleepornM, SchmidtP, et al. (2012) Gross karyotypic and phenotypic alterations among different progenies of the Candida glabrata CBS138/ATCC2001 reference strain. PLoS One 7: e52218.

8. PolakovaS, BlumeC, ZarateJA, MentelM, Jorck-RambergD, et al. (2009) Formation of new chromosomes as a virulence mechanism in yeast Candida glabrata. Proc Natl Acad Sci U S A 106: 2688–2693.

9. MannPA, McNicholasPM, ChauAS, PatelR, MendrickC, et al. (2009) Impact of antifungal prophylaxis on colonization and azole susceptibility of Candida species. Antimicrob Agents Chemother 53: 5026–5034.

10. vanden BosscheH, MarichalP, OddsFC, Le JeuneL, CoeneMC (1992) Characterization of an azole-resistant Candida glabrata isolate. Antimicrob Agents Chemother 36: 2602–2610.

11. WoodmanZ, WilliamsonC (2009) HIV molecular epidemiology: transmission and adaptation to human populations. Curr Opin HIV AIDS 4: 247–252.

12. ZiebuhrW, OhlsenK, KarchH, KorhonenT, HackerJ (1999) Evolution of bacterial pathogenesis. Cell Mol Life Sci 56: 719–728.

13. MorschhauserJ, KohlerG, ZiebuhrW, Blum-OehlerG, DobrindtU, et al. (2000) Evolution of microbial pathogens. Philos Trans R Soc Lond B Biol Sci 355: 695–704.

14. MaurelliAT (2007) Black holes, antivirulence genes, and gene inactivation in the evolution of bacterial pathogens. FEMS Microbiol Lett 267: 1–8.

15. JacobsenID, BrunkeS, SeiderK, SchwarzmullerT, FironA, et al. (2010) Candida glabrata persistence in mice does not depend on host immunosuppression and is unaffected by fungal amino acid auxotrophy. Infect Immun 78: 1066–1077.

16. SeiderK, BrunkeS, SchildL, JablonowskiN, WilsonD, et al. (2011) The facultative intracellular pathogen Candida glabrata subverts macrophage cytokine production and phagolysosome maturation. J Immunol 187: 3072–3086.

17. KaurR, MaB, CormackBP (2007) A family of glycosylphosphatidylinositol-linked aspartyl proteases is required for virulence of Candida glabrata. Proc Natl Acad Sci U S A 104: 7628–7633.

18. RoetzerA, GratzN, KovarikP, SchullerC (2010) Autophagy supports Candida glabrata survival during phagocytosis. Cell Microbiol 12: 199–216.

19. OttoV, HowardDH (1976) Further studies on the intracellular behavior of Torulopsis glabrata. Infect Immun 14: 433–438.

20. SteenbergenJN, NosanchukJD, MalliarisSD, CasadevallA (2003) Cryptococcus neoformans virulence is enhanced after growth in the genetically malleable host Dictyostelium discoideum. Infect Immun 71: 4862–4872.

21. CasadevallA (2008) Evolution of intracellular pathogens. Annu Rev Microbiol 62: 19–33.

22. MagditchDA, LiuTB, XueC, IdnurmA (2012) DNA mutations mediate microevolution between host-adapted forms of the pathogenic fungus Cryptococcus neoformans. PLoS Pathog 8: e1002936.

23. EnsmingerAW, YassinY, MironA, IsbergRR (2012) Experimental evolution of Legionella pneumophila in mouse macrophages leads to strains with altered determinants of environmental survival. PLoS Pathog 8: e1002731.

24. MiskinyteM, SousaA, RamiroRS, de SousaJA, KotlinowskiJ, et al. (2013) The Genetic Basis of Escherichia coli Pathoadaptation to Macrophages. PLoS Pathog 9: e1003802.

25. LachkeSA, JolyS, DanielsK, SollDR (2002) Phenotypic switching and filamentation in Candida glabrata. Microbiology 148: 2661–2674.

26. SchwarzmüllerT, MaB, HillerE, IstelF, TschernerM, et al. (2014) Systematic Phenotyping of a Large-Scale Candida glabrata Deletion Collection Reveals Novel Antifungal Tolerance Genes. PLoS Pathog 10: e1004211.

27. JacobsenID, GrosseK, BerndtA, HubeB (2011) Pathogenesis of Candida albicans infections in the alternative chorio-allantoic membrane chicken embryo model resembles systemic murine infections. PLoS One 6: e19741.

28. DujonB, ShermanD, FischerG, DurrensP, CasaregolaS, et al. (2004) Genome evolution in yeasts. Nature 430: 35–44.

29. De Las PenasA, PanSJ, CastanoI, AlderJ, CreggR, et al. (2003) Virulence-related surface glycoproteins in the yeast pathogen Candida glabrata are encoded in subtelomeric clusters and subject to RAP1- and SIR-dependent transcriptional silencing. Genes Dev 17: 2245–2258.

30. ThierryA, DujonB, RichardGF (2010) Megasatellites: a new class of large tandem repeats discovered in the pathogenic yeast Candida glabrata. Cell Mol Life Sci 67: 671–676.

31. ShawJA, MolPC, BowersB, SilvermanSJ, ValdiviesoMH, et al. (1991) The function of chitin synthases 2 and 3 in the Saccharomyces cerevisiae cell cycle. J Cell Biol 114: 111–123.

32. NagahashiS, SudohM, OnoN, SawadaR, YamaguchiE, et al. (1995) Characterization of chitin synthase 2 of Saccharomyces cerevisiae. Implication of two highly conserved domains as possible catalytic sites. J Biol Chem 270: 13961–13967.

33. KamranM, CalcagnoAM, FindonH, BignellE, JonesMD, et al. (2004) Inactivation of transcription factor gene ACE2 in the fungal pathogen Candida glabrata results in hypervirulence. Eukaryot Cell 3: 546–552.

34. VazquezJA, PengG, SobelJD, Steele-MooreL, SchumanP, et al. (2001) Evolution of antifungal susceptibility among Candida species isolates recovered from human immunodeficiency virus-infected women receiving fluconazole prophylaxis. Clin Infect Dis 33: 1069–1075.

35. FerrariS, IscherF, CalabreseD, PosteraroB, SanguinettiM, et al. (2009) Gain of function mutations in CgPDR1 of Candida glabrata not only mediate antifungal resistance but also enhance virulence. PLoS Pathog 5: e1000268.

36. SeiderK, HeykenA, LuttichA, MiramonP, HubeB (2010) Interaction of pathogenic yeasts with phagocytes: survival, persistence and escape. Curr Opin Microbiol 13: 392–400.

37. BrielandJ, EssigD, JacksonC, FrankD, LoebenbergD, et al. (2001) Comparison of pathogenesis and host immune responses to Candida glabrata and Candida albicans in systemically infected immunocompetent mice. Infect Immun 69: 5046–5055.

38. JuJY, PolhamusC, MarrKA, HollandSM, BennettJE (2002) Efficacies of fluconazole, caspofungin, and amphotericin B in Candida glabrata-infected p47phox−/− knockout mice. Antimicrob Agents Chemother 46: 1240–1245.

39. BrockertPJ, LachkeSA, SrikanthaT, PujolC, GalaskR, et al. (2003) Phenotypic switching and mating type switching of Candida glabrata at sites of colonization. Infect Immun 71: 7109–7118.

40. LinCY, ChenYC, LoHJ, ChenKW, LiSY (2007) Assessment of Candida glabrata strain relatedness by pulsed-field gel electrophoresis and multilocus sequence typing. J Clin Microbiol 45: 2452–2459.

41. MullerH, ThierryA, CoppeeJY, GouyetteC, HennequinC, et al. (2009) Genomic polymorphism in the population of Candida glabrata: gene copy-number variation and chromosomal translocations. Fungal Genet Biol 46: 264–276.

42. ThierryA, BouchierC, DujonB, RichardGF (2008) Megasatellites: a peculiar class of giant minisatellites in genes involved in cell adhesion and pathogenicity in Candida glabrata. Nucleic Acids Res 36: 5970–5982.

43. LuettichA, BrunkeS, HubeB, JacobsenID (2013) Serial passaging of Candida albicans in systemic murine infection suggests that the wild type strain SC5314 is well adapted to the murine kidney. PLoS One In press.

44. NeilsonJB, IveyMH, BulmerGS (1978) Cryptococcus neoformans: pseudohyphal forms surviving culture with Acanthamoeba polyphaga. Infect Immun 20: 262–266.

45. FriesBC, GoldmanDL, CherniakR, JuR, CasadevallA (1999) Phenotypic switching in Cryptococcus neoformans results in changes in cellular morphology and glucuronoxylomannan structure. Infect Immun 67: 6076–6083.

46. MacCallumDM, FindonH, KennyCC, ButlerG, HaynesK, et al. (2006) Different consequences of ACE2 and SWI5 gene disruptions for virulence of pathogenic and nonpathogenic yeasts. Infect Immun 74: 5244–5248.

47. O'ConallainC, DoolinMT, TaggartC, ThorntonF, ButlerG (1999) Regulated nuclear localisation of the yeast transcription factor Ace2p controls expression of chitinase (CTS1) in Saccharomyces cerevisiae. Mol Gen Genet 262: 275–282.

48. DohrmannPR, ButlerG, TamaiK, DorlandS, GreeneJR, et al. (1992) Parallel pathways of gene regulation: homologous regulators SWI5 and ACE2 differentially control transcription of HO and chitinase. Genes Dev 6: 93–104.

49. LorenzMC, BenderJA, FinkGR (2004) Transcriptional response of Candida albicans upon internalization by macrophages. Eukaryot Cell 3: 1076–1087.

50. HummertS, HummertC, SchroterA, HubeB, SchusterS (2010) Game theoretical modelling of survival strategies of Candida albicans inside macrophages. J Theor Biol 264: 312–318.

51. CharlierC, NielsenK, DaouS, BrigitteM, ChretienF, et al. (2009) Evidence of a role for monocytes in dissemination and brain invasion by Cryptococcus neoformans. Infect Immun 77: 120–127.

52. MaffeiCM, MirelsLF, SobelRA, ClemonsKV, StevensDA (2004) Cytokine and inducible nitric oxide synthase mRNA expression during experimental murine cryptococcal meningoencephalitis. Infect Immun 72: 2338–2349.

53. CsankC, HaynesK (2000) Candida glabrata displays pseudohyphal growth. FEMS Microbiol Lett 189: 115–120.

54. VandeputteP, TronchinG, BergesT, HennequinC, ChabasseD, et al. (2007) Reduced susceptibility to polyenes associated with a missense mutation in the ERG6 gene in a clinical isolate of Candida glabrata with pseudohyphal growth. Antimicrob Agents Chemother 51: 982–990.

55. JacobsenID, GrosseK, SlesionaS, HubeB, BerndtA, et al. (2010) Embryonated eggs as an alternative infection model to investigate Aspergillus fumigatus virulence. Infect Immun 78: 2995–3006.

56. HoffmanCS, WinstonF (1987) A ten-minute DNA preparation from yeast efficiently releases autonomous plasmids for transformation of Escherichia coli. Gene 57: 267–272.

57. SchönianG, MeuselO, TietzHJ, MeyerW, GraserY, et al. (1993) Identification of clinical strains of Candida albicans by DNA fingerprinting with the polymerase chain reaction. Mycoses 36: 171–179.

58. Rosas-HernandezLL, Juarez-ReyesA, Arroyo-HelgueraOE, De Las PenasA, PanSJ, et al. (2008) yKu70/yKu80 and Rif1 regulate silencing differentially at telomeres in Candida glabrata. Eukaryot Cell 7: 2168–2178.

59. Sambrook J, Russell D (2001) Molecular Cloning - A Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.

60. OssowskiS, SchneebergerK, ClarkRM, LanzC, WarthmannN, et al. (2008) Sequencing of natural strains of Arabidopsis thaliana with short reads. Genome Res 18: 2024–2033.