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Chitin-Degrading Protein CBP49 Is a Key Virulence Factor in American Foulbrood of Honey Bees
American Foulbrood and its etiological agent, Paenibacillus larvae, pose a serious threat to global honey bee health. So far, molecular mechanisms of host-microbe interactions are poorly understood in this system and no key virulence factor for the entire species has been identified. Here, we demonstrate that P. larvae expresses a chitin-binding and -degrading protein PlCBP49 harboring one module that belongs to the auxiliary activity 10 (AA10) family of lytic polysaccharide monooxygenases (LPMOs). We provide evidence that PlCBP49 degrades chitin via a metal ion-dependent, oxidative mechanism, as already described for other members of the AA10 enzyme family. Using P. larvae mutants lacking PlCBP49 expression, we demonstrate that PlCBP49 is crucial for the degradation of the chitin-rich peritrophic matrix, a key step in pathogenesis of P. larvae infections. In the absence of PlCBP49 expression the peritrophic matrix remained nearly intact and about 95% of the infected larvae survived infection. This clearly indicated that PlCBP49 is a key virulence factor of P. larvae. These results constitute important progress in our understanding of both P. larvae pathogenesis and the biological role of LPMOs in entomopathogens. Furthermore, knowing PlCBP49 and its role in pathogenesis opens new possibilities to develop curative measures for this disease.
Vyšlo v časopise: Chitin-Degrading Protein CBP49 Is a Key Virulence Factor in American Foulbrood of Honey Bees. PLoS Pathog 10(7): e32767. doi:10.1371/journal.ppat.1004284
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004284Souhrn
American Foulbrood and its etiological agent, Paenibacillus larvae, pose a serious threat to global honey bee health. So far, molecular mechanisms of host-microbe interactions are poorly understood in this system and no key virulence factor for the entire species has been identified. Here, we demonstrate that P. larvae expresses a chitin-binding and -degrading protein PlCBP49 harboring one module that belongs to the auxiliary activity 10 (AA10) family of lytic polysaccharide monooxygenases (LPMOs). We provide evidence that PlCBP49 degrades chitin via a metal ion-dependent, oxidative mechanism, as already described for other members of the AA10 enzyme family. Using P. larvae mutants lacking PlCBP49 expression, we demonstrate that PlCBP49 is crucial for the degradation of the chitin-rich peritrophic matrix, a key step in pathogenesis of P. larvae infections. In the absence of PlCBP49 expression the peritrophic matrix remained nearly intact and about 95% of the infected larvae survived infection. This clearly indicated that PlCBP49 is a key virulence factor of P. larvae. These results constitute important progress in our understanding of both P. larvae pathogenesis and the biological role of LPMOs in entomopathogens. Furthermore, knowing PlCBP49 and its role in pathogenesis opens new possibilities to develop curative measures for this disease.
Zdroje
1. RichardsAG, RichardsPA (1977) The peritrophic membranes of insects. Annu Rev Entomol 22 : 219–240.
2. TerraWR (2001) The origin and functions of the insect peritrophic membrane and peritrophic gel. Arch Insect Biochem Physiol 47 : 47–61.
3. HegedusD, ErlandsonM, GillottC, ToprakU (2009) New insights into peritrophic matrix synthesis, architecture, and function. Annu Rev Entomol 54 : 285–302.
4. CantarelBLea (2009) The Carbohydrate-Active EnZymes database (CAZy): an expert resource for glycogenomics. Nucleic Acids Res 37: D233–238.
5. Vaaje-KolstadG, HornSJ, van AaltenDMF, SynstadB, EijsinkVGH (2005) The non-catalytic chitin-binding protein CBP21 from Serratia marcescens is essential for chitin degradation. J Biol Chem 280 : 28492–28497.
6. GrimontPAD, GrimontF (1978) The genus Serratia. Annu Rev Microbiol 32 : 221–248.
7. JulianGS, BullaLAJr, SharpeES, AdamsGL (1973) Bacteria, spirochetes, and rickettsia as insecticides. Ann NY Acad Sci 217 : 65–75.
8. Vaaje-KolstadG, WesterengB, HornSJ, LiuZ, HZ, et al. (2010) An oxidative enzyme boosting the enzymatic conversion of recalcitrant polysaccharides. Science 330 : 219–222.
9. Vaaje-KolstadG, BøhleLA, GåseidnesS, DalhusB, BjøråsM, et al. (2012) Characterization of the chitinolytic machinery of Enterococcus faecalis V583 and high-resolution structure of its oxidative CBM33 enzyme. J Mol Biol 416 : 239–254.
10. AachmannFL, SørlieM, Skjåk-BrækG, EijsinkVGH, Vaaje-KolstadG (2012) NMR structure of a lytic polysaccharide monooxygenase provides insight into copper binding, protein dynamics, and substrate interactions. Proc Natl Acad Sci USA 109 : 18779–18784.
11. BolognesiR, TerraWR, FerreiraC (2008) Peritrophic membrane role in enhancing digestive efficiency:Theoretical and experimental models. J Insect Physiol 54 : 1413–1422.
12. BhattacharyaD, NagpureA, GuptaRK (2007) Bacterial Chitinases: Properties and Potential. Crit Rev Biotechnol 27 : 21–28.
13. GenerschE, ForsgrenE, PentikäinenJ, AshiralievaA, RauchS, et al. (2006) Reclassification of Paenibacillus larvae subsp. pulvifaciens and Paenibacillus larvae subsp. larvae as Paenibacillus larvae without subspecies differentiation. Int J Syst Evol Microbiol 56 : 501–511.
14. GenerschE (2007) Paenibacillus larvae and American foulbrood in honeybees. Berl Münch Tierärztl Wschr 120 : 26–33.
15. GenerschE (2008) Paenibacillus larvae and American foulbrood – long since known and still surprising. J Verbr Lebensm 3 : 429–434.
16. GenerschE (2010) American Foulbrood in honeybees and its causative agent, Paenibacillus larvae. J Invertebr Pathol 103: S10–S19.
17. DavidsonEW (1970) Ultrastructure of perithrophic membrane development in larvae of the worker honey bee (Apis mellifera). J Invertebr Pathol 15 : 451–454.
18. DavidsonEW (1973) Ultrastructure of American foulbrood disease pathogenesis in larvae of the worker honey bee, Apis mellifera. J Invertebr Pathol 21 : 53–61.
19. Garcia-GonzalezE, GenerschE (2013) Honey bee larval peritrophic matrix degradation during infection with Paenibacillus larvae, the aetiological agent of American foulbrood of honey bees, is a key step in pathogenesis. Environ Microbiol 15 : 2894–2901.
20. YueD, NordhoffM, WielerLH, GenerschE (2008) Fluorescence in situ-hybridization (FISH) analysis of the interactions between honeybee larvae and Paenibacillus larvae, the causative agent of American foulbrood of honeybees (Apis mellifera). Environ Microbiol 10 : 1612–1620.
21. HolstEC (1946) A simple field test for American foulbrood. Amer Bee 86 : 34.
22. HolstEC, SturtevantAP (1940) Relation of proteolytic enzymes to phase of life cycle of Bacillus larvae, and two new culture media for this organism. J Bacteriol 40 : 723–731.
23. DancerBN, ChantawannakulP (1997) The proteases of American foulbrood scales. J Invertebr Pathol 70 : 79–87.
24. AntúnezK, AnidoM, SchlappG, EvansJD, ZuninoP (2009) Characterization of secreted proteases of Paenibacillus larvae, potential virulence factors involved in honeybee larval infection. J Invertebr Pathol 102 : 129–132.
25. AntunezK, ArredondoD, AnidoM, ZuninoP (2011) Metalloprotease production by Paenibacillus larvae during the infection of honeybee larvae. Microbiology SGM 157 : 1474–1480.
26. DjukicM, BrzuszkiewiczE, FünfhausA, VossJ, GollnowK, et al. (2014) How to kill the honey bee larva: Genomic potential and virulence mechanisms of Paenibacillus larvae. PLoS ONE 9: e90914.
27. QinX, EvansJD, AronsteinKA, MurrayKD, WeinstockGM (2006) Genome sequence of the honey bee pathogens Paenibacillus larvae and Ascosphaera apis. Insect Mol Biol 15 : 715–718.
28. AltschulSF, MaddenTL, SchäfferAA, ZhangJ, ZhangZ, et al. (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25 : 3389–33402.
29. ManjeetK, PurushothamP, NeerajaC, PodileAR (2013) Bacterial chitin binding proteins show differential substrate binding and synergy with chitinases. Microbiol Res 168 : 461–468.
30. MehmoodMA, XiaoX, HafeezFY, GaiYB, WangFP (2011) Molecular characterization of the modular chitin binding protein Cbp50 from Bacillus thuringiensis serovar konkukian. Anton Leeuw Int J G 100 : 445–453.
31. MoserF, IrwinD, ChenS, WilsonDB (2008) Regulation and characterization of Thermobifida fusca carbohydrate-binding module proteins E7 and E8. Biotechnol Bioeng 100 : 1066–1077.
32. RaoFV, AndersenOA, VoraKA, DeMartinoJA, van AaltenDMF (2005) Methylxanthine drugs are chitinase inhibitors: Investigation of inhibition and binding modes. Chem Biol 12 : 973–980.
33. BamrickJ-F (1967) Resistance to American foulbrood in honey bees. VI. Spore germination in larvae of different ages. J Invertebr Pathol 9 : 30–34.
34. JaeckelS (1930) Zur pathologischen Anatomie der Biene Apis mellifica L. während der Metamorphose bei bösartiger Faulbrut. Archiv für Bienenkunde 11 : 41–92.
35. BamrickJ-F (1964) Resistance to American foulbrood in honey bees. V. Comparative pathogenesis in resistant and susceptible larvae. J Insect Pathol 6 : 284–304.
36. Shapiro-IlanDI, FuxaJR, LaceyLA, OnstadDW, KayaHK (2005) Definitions of pathogenicity and virulence in invertebrate pathology. J Invertebr Pathol 88 : 1–7.
37. FlygC, KenneK, BomanHG (1980) Insect pathogenic properties of Serratia marcescens: Phageresistant mutants with a decreased resistance to Cecropia immunity and a decreased virulence to Drosophila. J Gen Microbiol 120 : 173–181.
38. HejaziA, FalkinerFR (1997) Serratia marcescens. J Med Microbiol 46 : 903–912.
39. KuraishiT, BinggeliO, OpotaO, BuchonN, LemaitreB (2011) Genetic evidence for a protective role of the peritrophic matrix against intestinal bacterial infection in Drosophila melanogaster. Proc Natl Acad Sci USA 108 : 15966–15971.
40. NakjangS, NdehDA, WipatA, BolamDN, HirtRP (2012) A novel extracellular metallopeptidase domain shared by animal host-assocaited mutualistic and pathogenic microbes. PLoS ONE 7: e30287.
41. HayakawaT, ShitomiY, MiyamotoK, HoriH (2004) GalNAc pretreatment inhibits trapping of Bacillus thuringiensis Cry1Ac on the peritrophic membrane of Bombyx mori. FEBS Lett 576 : 331–335.
42. FünfhausA, AshiralievaA, BorrissR, GenerschE (2009) Use of suppression subtractive hybridization to identify genetic differences between differentially virulent genotypes of Paenibacillus larvae, the etiological agent of American Foulbrood of honeybees. Environ Microbiol Rep 1 : 240–250.
43. FünfhausA, PoppingaL, GenerschE (2013) Identification and characterization of two novel toxins expressed by the lethal honey bee pathogen Paenibacillus larvae, the causative agent of American foulbrood. Environ Microbiol 15 : 2951–2965.
44. BarthH, AktoriesK, PopoffMR, StilesBG (2004) Binary bacterial toxins: biochemistry, biology, and applications of common Clostridium and Bacillus proteins. Microbiol Mol Biol Rev 68 : 373–402.
45. HolbournKP, ShoneCC, AcharyaKR (2006) A family of killer toxins: Exploring the mechanism of ADP-ribosylating toxins. FEBS J 273 : 4579–4593.
46. PoppingaL, JaneschB, FünfhausA, SekotG, Garcia-GonzalezE, et al. (2012) Identification and functional analysis of the S-layer protein SplA of Paenibacillus larvae, the causative agent of American Foulbrood of honey bees. PLoS Path 8: e1002716.
47. GenerschE, OttenC (2003) The use of repetitive element PCR fingerprinting (rep-PCR) for genetic subtyping of German field isolates of Paenibacillus larvae subsp. larvae. Apidologie 34 : 195–206.
48. GenerschE, AshiralievaA, FriesI (2005) Strain - and genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, the causative agent of American foulbrood disease in honey bees. Appl Environ Microbiol 71 : 7551–7555.
49. RauchS, AshiralievaA, HedtkeK, GenerschE (2009) Negative correlation between individual-insect-level virulence and colony-level virulence of Paenibacillus larvae, the etiological agent of American foulbrood of honeybees. Appl Environ Microbiol 75 : 3344–3347.
50. FünfhausA, GenerschE (2012) Proteome analysis of Paenibacillus larvae reveals the existence of a putative S-layer protein. Environ Microbiol Rep 4 : 194–202.
51. NeuendorfS, HedtkeK, TangenG, GenerschE (2004) Biochemical characterization of different genotypes of Paenibacillus larvae subsp. larvae, a honey bee bacterial pathogen. Microbiology 150 : 2381–2390.
52. DingmanDW, StahlyDP (1983) Medium promoting sporulation of Bacillus larvae and metabolism of medium components. Appl Environ Microbiol 46 : 860–869.
53. ZarschlerK, JaneschB, PabstM, AltmannF, MessnerP, et al. (2010) Protein tyrosine O-glycosylation—A rather unexplored prokaryotic glycosylation system. Glycobiology 20 : 787–798.
54. LaemmliUK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 : 680–685.
55. TrudelJ, AsselinA (1989) Detection of chitinase activity after polyacrylamide gel electrophoresis. Anal Biochem 178 : 362–366.
56. ZarschlerK, JaneschB, ZayniS, SchäfferC, MessnerP (2009) Construction of a gene knockout system for application in Paenibacillus alvei CCM 2051T, exemplified by the S-layer glycan biosynthesis initiation enzyme WsfP. Appl Environ Microbiol 75 : 3077–3085.
57. MurrayKD, AronsteinKA (2008) Transformation of the Gram-positive honey bee pathogen, Paenibacillus larvae, by electroporation. J Microbiol Meth 75 : 325–328.
58. PoppingaL, GenerschE (2012) Heterologous expression of green fluorescent protein in Paenibacillus larvae, the causative agent of American Foulbrood of honey bees. J Appl Microbiol 112 : 430–435.
59. JakubowskaAK, CacciaS, GordonKH, FerréJ, HerreroS (2010) Downregulation of a chitin deacetylase-like protein in response to baculovirus infection and its application for improving baculovirus infectivity. J Virol 84 : 2547–2555.
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