#PAGE_PARAMS# #ADS_HEAD_SCRIPTS# #MICRODATA#

Domain Shuffling in a Sensor Protein Contributed to the Evolution of Insect Pathogenicity in Plant-Beneficial


Pseudomonas protegens is a biocontrol rhizobacterium with a plant-beneficial and an insect pathogenic lifestyle, but it is not understood how the organism switches between the two states. Here, we focus on understanding the function and possible evolution of a molecular sensor that enables P. protegens to detect the insect environment and produce a potent insecticidal toxin specifically during insect infection but not on roots. By using quantitative single cell microscopy and mutant analysis, we provide evidence that the sensor histidine kinase FitF is a key regulator of insecticidal toxin production. Our experimental data and bioinformatic analyses indicate that FitF shares a sensing domain with DctB, a histidine kinase regulating carbon uptake in Proteobacteria. This suggested that FitF has acquired its specificity through domain shuffling from a common ancestor. We constructed a chimeric DctB-FitF protein and showed that it is indeed functional in regulating toxin expression in P. protegens. The shuffling event and subsequent adaptive modifications of the recruited sensor domain were critical for the microorganism to express its potent insect toxin in the observed host-specific manner. Inhibition of the FitF sensor during root colonization could explain the mechanism by which P. protegens differentiates between the plant and insect host. Our study establishes FitF of P. protegens as a prime model for molecular evolution of sensor proteins and bacterial pathogenicity.


Vyšlo v časopise: Domain Shuffling in a Sensor Protein Contributed to the Evolution of Insect Pathogenicity in Plant-Beneficial. PLoS Pathog 10(2): e32767. doi:10.1371/journal.ppat.1003964
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003964

Souhrn

Pseudomonas protegens is a biocontrol rhizobacterium with a plant-beneficial and an insect pathogenic lifestyle, but it is not understood how the organism switches between the two states. Here, we focus on understanding the function and possible evolution of a molecular sensor that enables P. protegens to detect the insect environment and produce a potent insecticidal toxin specifically during insect infection but not on roots. By using quantitative single cell microscopy and mutant analysis, we provide evidence that the sensor histidine kinase FitF is a key regulator of insecticidal toxin production. Our experimental data and bioinformatic analyses indicate that FitF shares a sensing domain with DctB, a histidine kinase regulating carbon uptake in Proteobacteria. This suggested that FitF has acquired its specificity through domain shuffling from a common ancestor. We constructed a chimeric DctB-FitF protein and showed that it is indeed functional in regulating toxin expression in P. protegens. The shuffling event and subsequent adaptive modifications of the recruited sensor domain were critical for the microorganism to express its potent insect toxin in the observed host-specific manner. Inhibition of the FitF sensor during root colonization could explain the mechanism by which P. protegens differentiates between the plant and insect host. Our study establishes FitF of P. protegens as a prime model for molecular evolution of sensor proteins and bacterial pathogenicity.


Zdroje

1. HaasD, DéfagoG (2005) Biological control of soil-borne pathogens by fluorescent pseudomonads. Nat Rev Microbiol 3: 307–319.

2. RametteA, FrapolliM, Fischer-Le SauxM, GruffazC, MeyerJM, et al. (2011) Pseudomonas protegens sp nov., widespread plant-protecting bacteria producing the biocontrol compounds 2,4-diacetylphloroglucinol and pyoluteorin. Syst Appl Microbiol 34: 180–188.

3. KupferschmiedP, MaurhoferM, KeelC (2013) Promise for plant pest control: root-associated pseudomonads with insecticidal activities. Front Plant Sci 4: 287.

4. LoperJE, HassanKA, MavrodiDV, DavisEW, LimCK, et al. (2012) Comparative genomics of plant-associated Pseudomonas spp.: insights into diversity and inheritance of traits involved in multitrophic nteractions. PLoS Genet 8: e1002784.

5. RuffnerB, Péchy-TarrM, RyffelF, HoeggerP, ObristC, et al. (2013) Oral insecticidal activity of plant-associated pseudomonads. Environ Microbiol 15: 751–763.

6. Péchy-TarrM, BruckDJ, MaurhoferM, FischerE, KeelC (2008) Molecular analysis of a novel gene cluster encoding an insect toxin in plant-associated strains of Pseudomonas fluorescens. Environ Microbiol 10: 2368–2386.

7. Péchy-TarrM, BorelN, KupferschmiedP, TurnerV, BinggeliO, et al. (2013) Control and host-dependent activation of insect toxin expression in a root-associated biocontrol pseudomonad. Environ Microbiol 15: 736–750.

8. GaoR, StockAM (2009) Biological insights from structures of two-component proteins. Annu Rev Microbiol 63: 133–154.

9. KrellT, LacalJ, BuschA, Silva-JimenezH, GuazzaroniME, et al. (2010) Bacterial sensor kinases: diversity in the recognition of environmental signals. Annu Rev Microbiol 64: 539–559.

10. AlmE, HuangK, ArkinA (2006) The evolution of two-component systems in bacteria reveals different strategies for niche adaptation. PLoS Comput Biol 2: e143.

11. GalperinMY (2005) A census of membrane-bound and intracellular signal transduction proteins in bacteria: bacterial IQ, extroverts and introverts. BMC Microbiol 5: 35.

12. CapraEJ, LaubMT (2012) Evolution of two-component signal transduction systems. Annu Rev Microbiol 66: 325–347.

13. RaghavanV, GroismanEA (2010) Orphan and hybrid two-component system proteins in health and disease. Curr Opin Microbiol 13: 226–231.

14. JungK, FriedL, BehrS, HeermannR (2012) Histidine kinases and response regulators in networks. Curr Opin Microbiol 15: 118–124.

15. ScheuPD, KimOB, GriesingerC, UndenG (2010) Sensing by the membrane-bound sensor kinase DcuS: exogenous versus endogenous sensing of C(4)-dicarboxylates in bacteria. Future Microbiol 5: 1383–1402.

16. CheungJ, HendricksonWA (2008) Crystal structures of C-4-dicarboxylate ligand complexes with sensor domains of histidine kinases DcuS and DctB. J Biol Chem 283: 30256–30265.

17. ZhouYF, NanBY, NanJ, MaQJ, PanjikarS, et al. (2008) C(4)-dicarboxylates sensing mechanism revealed by the crystal structures of DctB sensor domain. J Mol Biol 383: 49–61.

18. CheungJ, HendricksonWA (2010) Sensor domains of two-component regulatory systems. Curr Opin Microbiol 13: 116–123.

19. ZhangW, ShiL (2005) Distribution and evolution of multiple-step phosphorelay in prokaryotes: lateral domain recruitment involved in the formation of hybrid-type histidine kinases. Microbiology 151: 2159–2173.

20. ChangC, TesarC, GuM, BabniggG, JoachimiakA, et al. (2010) Extracytoplasmic PAS-like domains are common in signal transduction proteins. J Bacteriol 192: 1156–1159.

21. HenryJT, CrossonS (2011) Ligand-binding PAS domains in a genomic, cellular, and structural context. Annu Rev Microbiol 65: 261–286.

22. StephensonK, HochJA (2002) Evolution of signalling in the sporulation phosphorelay. Mol Microbiol 46: 297–304.

23. RiechmannL, WinterG (2000) Novel folded protein domains generated by combinatorial shuffling of polypeptide segments. Proc Natl Acad Sci U S A 97: 10068–10073.

24. MöglichA, AyersRA, MoffatK (2010) Addition at the molecular level: signal integration in designed Per-ARNT-Sim receptor proteins. J Mol Biol 400: 477–486.

25. ChecaSK, ZurbriggenMD, SonciniFC (2012) Bacterial signaling systems as platforms for rational design of new generations of biosensors. Curr Opin Biotechnol 23: 766–772.

26. GraceTD (1962) Establishment of four strains of cells from insect tissues grown in vitro. Nature 195: 788–789.

27. ShenX, HuH, PengH, WangW, ZhangX (2013) Comparative genomic analysis of four representative plant growth-promoting rhizobacteria in Pseudomonas. BMC Genomics 14: 271.

28. NanBY, LiuX, ZhouYF, LiuJW, ZhangL, et al. (2010) From signal perception to signal transduction: ligand-induced dimeric switch of DctB sensory domain in solution. Mol Microbiol 75: 1484–1494.

29. ZhangZ, HendricksonWA (2010) Structural characterization of the predominant family of histidine kinase sensor domains. J Mol Biol 400: 335–353.

30. EtzkornM, KneuperH, DunnwaldP, VijayanV, KramerJ, et al. (2008) Plasticity of the PAS domain and a potential role for signal transduction in the histidine kinase DcuS. Nat Struct Mol Biol 15: 1031–1039.

31. ValentiniM, StorelliN, LapougeK (2011) Identification of C(4)-dicarboxylate transport systems in Pseudomonas aeruginosa PAO1. J Bacteriol 193: 4307–4316.

32. PetrovaOE, SauerK (2009) A novel signaling network essential for regulating Pseudomonas aeruginosa biofilm development. PLoS Pathog 5: e1000668.

33. QianW, HanZJ, HeC (2008) Two-component signal transduction systems of Xanthomonas spp.: a lesson from genomics. Mol Plant Microbe Interact 21: 151–161.

34. Sambrook J, Russel DW (2001) Molecular Cloning: A Laboratory Manual. Cold Spring Harbor (New York): Cold Spring Harbor Laboratory Press.

35. Schnider-KeelU, SeematterA, MaurhoferM, BlumerC, DuffyB, et al. (2000) Autoinduction of 2,4-diacetylphloroglucinol biosynthesis in the biocontrol agent Pseudomonas fluorescens CHA0 and repression by the bacterial metabolites salicylate and pyoluteorin. J Bacteriol 182: 1215–1225.

36. Défago G, Haas D (1990) Pseudomonads as antagonists of soilborne plant pathogens: modes of action and genetic analysis. In: Bollag JM, Stotzky G, editors. Soil Biochemistry. New York, USA: Marcel Dekker. pp. 249–291.

37. Martinez-GarciaE, de LorenzoV (2011) Engineering multiple genomic deletions in Gram-negative bacteria: analysis of the multi-resistant antibiotic profile of Pseudomonas putida KT2440. Environ Microbiol 13: 2702–2716.

38. BaehlerE, BottiglieriM, Pechy-TarrM, MaurhoferM, KeelC (2005) Use of green fluorescent protein-based reporters to monitor balanced production of antifungal compounds in the biocontrol agent Pseudomonas fluorescens CHA0. J Appl Microbiol 99: 24–38.

39. Sharifi-TehraniA, ZalaM, NatschA, Moenne-LoccozY, DefagoG (1998) Biocontrol of soil-borne fungal plant diseases by 2,4-diacetylphloroglucinol-producing fluorescent pseudomonads with different restriction profiles of amplified 16S rDNA. Eur J Plant Pathol 104: 631–643.

40. AltschulSF, MaddenTL, SchafferAA, ZhangJ, ZhangZ, et al. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389–3402.

41. CserzoM, WallinE, SimonI, von HeijneG, ElofssonA (1997) Prediction of transmembrane alpha-helices in prokaryotic membrane proteins: the dense alignment surface method. Protein Eng 10: 673–676.

42. Marchler-BauerA, LuS, AndersonJB, ChitsazF, DerbyshireMK, et al. (2011) CDD: a Conserved Domain Database for the functional annotation of proteins. Nucleic Acids Res 39: D225–229.

43. SchultzJ, MilpetzF, BorkP, PontingCP (1998) SMART, a simple modular architecture research tool: identification of signaling domains. Proc Natl Acad Sci U S A 95: 5857–5864.

44. TamuraK, PetersonD, PetersonN, StecherG, NeiM, et al. (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28: 2731–2739.

45. FrickeyT, LupasA (2004) CLANS: a Java application for visualizing protein families based on pairwise similarity. Bioinformatics 20: 3702–3704.

46. KrämerJ, FischerJD, ZientzE, VijayanV, GriesingerC, et al. (2007) Citrate sensing by the C-4-dicarboxylate/citrate sensor kinase DcuS of Escherichia coli: Binding site and conversion of DcuS to a C-4-dicarboxylate- or citrate-specific sensor. J Bacteriol 189: 4290–4298.

47. LambertC, LeonardN, De BolleX, DepiereuxE (2002) ESyPred3D: Prediction of proteins 3D structures. Bioinformatics 18: 1250–1256.

48. RoyA, KucukuralA, ZhangY (2010) I-TASSER: a unified platform for automated protein structure and function prediction. Nat Protoc 5: 725–738.

49. WuS, ZhangY (2007) LOMETS: a local meta-threading-server for protein structure prediction. Nucleic Acids Res 35: 3375–3382.

50. KelleyLA, SternbergMJ (2009) Protein structure prediction on the Web: a case study using the Phyre server. Nat Protoc 4: 363–371.

51. ArnoldK, BordoliL, KoppJ, SchwedeT (2006) The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics 22: 195–201.

Štítky
Hygiena a epidemiológia Infekčné lekárstvo Laboratórium

Článok vyšiel v časopise

PLOS Pathogens


2014 Číslo 2
Najčítanejšie tento týždeň
Najčítanejšie v tomto čísle
Kurzy

Zvýšte si kvalifikáciu online z pohodlia domova

Získaná hemofilie - Povědomí o nemoci a její diagnostika
nový kurz

Eozinofilní granulomatóza s polyangiitidou
Autori: doc. MUDr. Martina Doubková, Ph.D.

Všetky kurzy
Prihlásenie
Zabudnuté heslo

Zadajte e-mailovú adresu, s ktorou ste vytvárali účet. Budú Vám na ňu zasielané informácie k nastaveniu nového hesla.

Prihlásenie

Nemáte účet?  Registrujte sa

#ADS_BOTTOM_SCRIPTS#