Crystal structure of the effector AvrLm4-7 of Leptosphaeria maculans reveals insights into its translocation into plant cells and recognition by resistance proteins

Plant Journal

Volumn 83, Issue 4, 2015, Pages 610-624

  Blondeau, Karine ^a   Blaise, Françoise ^b   Graille, Marc ^a   Kale, Shiv D ^c   Linglin, Juliette ^b   Ollivier, Bénédicte ^b   Labarde, Audrey ^a   Lazar, Noureddine ^a   Daverdin, Guillaume ^b   Balesdent, Marie Hélène ^b   Choi, Danielle H Y ^c   Tyler, Brett M ^c,d   Rouxel, Thierry ^b   Van Tilbeurgh, Herman ^a   Fudal, Isabelle ^b  

^a UNIVERSITÉ PARIS SACLAY   (France)

^b INRA Institut National de la Recherche Agronomique   (France)

^c VIRGINIA BIOINFORMATICS INSTITUTE   (United States)

^d OREGON STATE UNIVERSITY   (United States)

Author keywords

3D structure; avirulence protein; Brassica napus; effector; Leptosphaeria maculans; plant cell translocation; plant disease resistance

Indexed keywords

AMINO ACIDS; COVALENT BONDS; GENES; OILSEEDS; PLANT CELL CULTURE; PLANTS (BOTANY); PROTEINS; SULFUR COMPOUNDS;

3D STRUCTURE; AVIRULENCE PROTEINS; BRASSICA NAPUS; EFFECTOR; LEPTOSPHAERIA MACULANS; PLANT CELLS; PLANT DISEASE RESISTANCES;

CRYSTAL STRUCTURE;

BRASSICA NAPUS; LEPTOSPHAERIA MACULANS; NICOTIANA BENTHAMIANA; NICOTIANA TABACUM; PICHIA PASTORIS;

FUNGAL PROTEIN;

ASCOMYCETES; GENETICS; METABOLISM; MICROBIOLOGY; PATHOGENICITY; PLANT DISEASE; RAPESEED; VIRULENCE;

ASCOMYCOTA; BRASSICA NAPUS; FUNGAL PROTEINS; PLANT DISEASES; VIRULENCE;

EID: 84938993930     PISSN: 09607412     EISSN: 1365313X     Source Type: Journal    
DOI: 10.1111/tpj.12913     Document Type: Article

References (58)

1. Ansan-Melayah, D., Balesdent, M.H., Buée, M., and, Rouxel, T., (1995) Genetic characterization of AvrLm1, the first avirulence gene of Leptosphaeria maculans. Phytopathology, 85, 1525-1529.
2. Baker, N.A., Sept, D., Joseph, S., Holst, M.J., and, McCammon, J.A., (2001) Electrostatics of nanosystems: application to microtubules and the ribosome. Proc. Natl Acad. Sci. USA, 98, 10037-10041.
3. Balesdent, M.H., Attard, A., Ansan-Melayah, D., Delourme, R., Renard, M., and, Rouxel, T., (2001) Genetic control and host range of avirulence toward Brassica napus cultivars Quinta and Jet Neuf in Leptosphaeria maculans. Phytopathology, 91, 70-76.
4. Balesdent, M.H., Attard, A., Kuhn, M.L., and, Rouxel, T., (2002) New avirulence genes in the phytopathogenic fungus Leptosphaeria maculans. Phytopathology, 92, 1122-1133.
5. Balesdent, M.H., Louvard, K., Pinochet, X., and, Rouxel, T., (2006) A large-scale survey of Leptosphaeria maculans occuring on oilseed rape in France. Eur. J. Plant Pathol. 114, 53-65.
6. Balesdent, M.H., Fudal, I., Ollivier, B., Bally, P., Grandaubert, J., Eber, F., Chèvre, A.M., Leflon, M., and, Rouxel, T., (2013) The dispensable chromosome of Leptosphaeria maculans shelters an effector gene conferring avirulence toward Brassica rapa. New Phytol. 198, 887-898.
7. Boller, T., and, Felix, G., (2009) A renaissance of elicitors: perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Annu. Rev. Plant Biol. 60, 379-406.
8. Boller, T., and, He, S.Y., (2009) Innate immunity in plants: an arms race between pattern recognition receptors in plants and effectors in microbial pathogens. Science, 324, 742-744.
9. Boutemy, L.S., King, S.R., Win, J., Hughes, R.K., Clarke, T.A., Blumenschein, T.M., Kamoun, S., and, Banfield, M.J., (2011) Structures of Phytophthora RXLR effector proteins: a conserved but adaptable fold underpins functional diversity. J. Biol. Chem. 286, 35834-35842.
10. Chisholm, S.T., Coaker, G., Day, B., and, Staskawicz, B.J., (2006) Host-microbe interactions: shaping the evolution of the plant immune response. Cell, 124, 803-814.
11. Chou, S., Krasileva, K.V., Holton, J.M., Steinbrenner, A.D., Alber, A., and, Staskawicz, B.J., (2011) Hyaloperonospora arabidopsidis ATR1 effector is a repeat protein with distributed recognition surfaces. Proc. Natl Acad. Sci. USA, 108, 13323-13328.
12. Dangl, J.L., and, McDowell, J.M., (2006) Two modes of pathogen recognition by plants. Proc. Natl Acad. Sci. USA, 103, 8575-8576.
13. Daverdin, G., Rouxel, T., Gout, L., Aubertot, J.N., Fudal, I., Meyer, M., Parlange, F., Carpezat, J., and, Balesdent, M.H., (2012) Genome structure and reproductive behaviour influence the evolutionary potential of a fungal phytopathogen. PLoS Pathog. 8, e1003020.
14. De Jonge, R., Bolton, M.D., and, Thomma, B.P.H.J., (2011) How filamentous pathogens co-opt plants: the ins and outs of fungal effectors. Curr. Opin. Plant Biol. 14, 1-7.
15. Dodds, P.N., Rafiqi, M., Gan, P.H., Hardham, A.R., Jones, D.A., and, Ellis, J.G., (2009) Effectors of biotrophic fungi and oomycetes: pathogenicity factors and triggers of host resistance. New Phytol. 183, 993-1000.
16. Dou, D., Kale, S.D., Wang, X., Jiang, R.H.Y., Bruce, N.A., Arredondo, F.D., Zhang, X., and, Tyler, B.M., (2008a) RXLR-mediated entry of Phytophthora sojae effector Avr1b into soybean cells does not require pathogen-encoded machinery. Plant Cell, 20, 1930-1947.
17. Dou, D., Kale, S.D., Wang, X., et al. (2008b) Conserved C-terminal motifs required for avirulence and suppression of cell death by Phytophthora sojae effector Avr1b. Plant Cell, 20, 1118-1133.
18. Fudal, I., Ross, S., Gout, L., Blaise, F., Kuhn, M.L., Eckert, M.R., Cattolico, L., Bernard-Samain, S., Balesdent, M.H., and, Rouxel, T., (2007) Heterochromatin-like regions as ecological niches for avirulence genes in the Leptosphaeria maculans genome: map-based cloning of AvrLm6. Mol. Plant Microbe Interact. 20, 459-470.
19. Gan, P.H.P., Rafiqi, M., Ellis, J.G., Jones, D.A., Hardham, A.R., and, Dodds, P.N., (2010) Lipid binding activities of flax rust AvrM and AvrL567 effectors. Plant Signaling and Behaviour, 5, 1272-1775.
20. Ghanbarnia, K., Fudal, I., Larkan, N.J., Links, M.G., Balesdent, M.H., Profotova, B., Fernando, W.G., Rouxel, T., and, Borhan, M.H., (2014) Rapid identification of the Leptosphaeria maculans avirulence gene AvrLm2 using an intraspecific comparative genomics approach. Mol. Plant Pathol., 16, 699-709.
21. Gout, L., Fudal, I., Kuhn, M.L., Blaise, F., Eckert, M., Cattolico, L., Balesdent, M.H., and, Rouxel, T., (2006) Lost in the middle of nowhere: the AvrLm1 avirulence gene of the dothideomycete Leptosphaeria maculans. Mol. Microbiol. 60, 67-80.
22. Grouffaud, S., van West, P., Avrova, A.O., Birch, P.R., and, Whisson, S.C., (2008) Plasmodium falciparum and Hyaloperonospora parasitica effector translocation motifs are functional in Phytophthora infestans. Microbiology, 154, 3743-3751.
23. Houterman, P.M., Cornelissen, B.J.C., and, Rep, M., (2008) Suppression of plant resistance gene-based immunity by a fungal effector. PLoS Pathog. 4, e1000061.
24. Huang, Y.J., Li, Z.Q., Evans, N., Rouxel, T., Fitt, B.D.L., and, Balesdent, M.H., (2006) Fitness cost associated with loss of the AvrLm4 avirulence function in Leptosphaeria maculans (phoma stem canker of oilseed rape). Eur. J. Plant Pathol. 114, 77-89.
25. Huang, Y.J., Balesdent, M.H., Li, Z.Q., Evans, N., Rouxel, T., and, Fitt, B.D.L., (2010) Fitness cost of virulence differs between the AvrLm1 and AvrLm4 loci in Leptosphaeria maculans (phoma stem canker of oilseed rape). Eur. J. Plant Pathol. 126, 279-291.
26. Jones, J.D., and, Dangl, J.L., (2006) The plant immune system. Nature, 444, 323-329.
27. Kale, S.D., and, Tyler, B.M., (2011a) Entry of oomycete and fungal effectors into plant and animal cells. Cellular Microbiol. 13, 1839-1848.
28. Kale, S.D., and, Tyler, B.M., (2011b) Assaying effector function in planta using double-barreled particle bombardment. Methods Mol. Biol. 712, 153-172.
29. Kale, S.D., Gu, B., Capelluto, D.F.S., et al. (2010) External lipid PI-3-P mediates entry of eukaryotic pathogen effectors into plant and animal host cells. Cell, 142, 284-295.
30. Larkan, N.J., Lydiate, D.J., Parkin, I.A.P., Nelson, M.N., Epp, D.J., Cowling, W.A., Rimmer, S.R., and, Borhan, M.H., (2013) The Brassica napus blackleg resistance gene LepR3 encodes a receptor-like protein triggered by the Leptosphaeria maculans effector AvrLm1. New Phytol. 197, 595-605.
31. Larkan, N.J., Ma, L., and, Borhan, M.H., (2015) The Brassica napus receptor-like protein Rlm2 is encoded by a second allele of the LepR3/Rlm2 blackleg resistance locus. Plant Biotechnol. J. doi: 10.1111/pbi.12341. [Epub ahead of print]
32. Larsson, A.M., and, Jones, T.A., (2007) Selenomethionine labeling of recombinant proteins. Methods Mol. Biol. 389, 165-174.
33. Larsson, A.M., Ståhlberg, J., and, Jones, T.A., (2002) Preparation and crystallization of selenomethionyl dextranase from Penicillium minioluteum expressed in Pichia pastoris. Acta Crystallogr. D Biol. Crystallogr. 58, 346-348.
34. Lo Presti, L., Lanver, D., Schweizer, G., Tanaka, S., Liang, L., Tollot, M., Zuccaro, A., Reissmann, S., and, Kahmann, R., (2015) Fungal effectors and plant susceptibility. Annu. Rev. Plant Biol. 66, 513-545.
35. Parlange, F., Daverdin, G., Fudal, I., Kuhn, M.L., Balesdent, M.H., Blaise, F., Grezes-Besset, B., and, Rouxel, T., (2009) Leptosphaeria maculans avirulence gene AvrLm4-7 confers a dual recognition specificity by the Rlm4 and Rlm7 resistance genes of oilseed rape, and circumvents Rlm4-mediated recognition through a single amino acid change. Mol. Microbiol. 71, 851-863.
36. Pedersen, C., Loren, V., van Themaat, E., et al. (2012) Structure and evolution of barley powdery mildew effector candidates. BMC Genom. 13, 694.
37. Petre, B., and, Kamoun, S., (2014) How do filamentous pathogens deliver effector proteins into plant cells? PLoS Biol. 12, e1001801.
38. Plett, J.M., Kemppainen, M., Kale, S.D., Kohler, A., Legué, V., Brun, A., Tyler, B.M., Pardo, A.G., and, Martin, F., (2011) A secreted effector protein of Laccaria bicolor is required for symbiosis development. Curr. Biol. 21, 1197-1203.
39. Rafiqi, M., Gan, P.H.P., Ravensdale, M., Lawrence, G.J., Ellis, J.G., Jones, D.A., Hardham, A.R., and, Dodds, P.N., (2010) Internalization of flax rust avirulence proteins into flax and tobacco cells can occur in the absence of the pathogen. Plant Cell, 22, 2017-2032.
40. Robert, X., and, Gouet, P., (2014) Deciphering key features in protein structures with the new ENDscript server. Nucleic Acids Res. 42, W320-324; doi: 10.1093/nar/gku316.
41. Rouxel, T., Grandaubert, J., Hane, J.K., et al. (2011) Effectors diversification within compartments of the Leptosphaeria maculans genome affected by RIP mutations. Nat. Commun. 2, 202.
42. Sacristan, S., and, Garcia-Arenal, F., (2008) The evolution of virulence and pathogenicity in plant pathogen populations. Mol. Plant Pathol. 9, 369-384.
43. Sanchez-Vallet, A., Saleem-Batcha, R., Kombrink, A., Hansen, G., Valkenburg, D.J., Mesters, J.R., and, Thomma, B.P.H.J., (2013) Fungal effector Ecp6 outcompetes host immune receptor for chitin binding through intrachain LysM dimerization. eLife, 2, e00790.
44. Sarma, G.N., Manning, V.A., Ciuffetti, L.M., and, Karplus, P.A., (2005) Structure of Ptr ToxA: an RGD-containing host-selective toxin from Pyrenophora tritici-repentis. Plant Cell, 17, 3190-3202.
45. Stassen, J.H.M., and, van den Ackerveken, G., (2011) How do oomycete effectors interfere with plant life? Curr. Opin. Plant Biol. 14, 1-8.
46. Stergiopoulos, I., and, de Wit, P.J.G.M., (2009) Fungal effector proteins. Annu. Rev. Phytopathol. 47, 233-263.
47. Sun, F., Kale, S.D., Azurmendi, H.F., Li, D., Tyler, B.M., and, Capelluto, D.G., (2013) Structural basis for interactions of Phytophthora sojae RXLR effector Avh5 with Phosphatidylinositol 3-phosphate and for host cell entry. Mol. Plant Microbe Interact. 26, 330-344.
48. Tyler, B.M., Kale, S.D., Wang, Q., et al. (2013) Microbe-independent entry of oomycete RxLR effectors and fungal RxLR-like effectors into plant and animal cells is specific and reproducible. Mol. Plant Microbe Interact. 26, 611-616.
49. Van de Wouw, A.P., Lowe, R.G., Elliott, C.E., Dubois, D.J., and, Howlett, B.J., (2013) An avirulence gene, AvrLmJ1, from the blackleg fungus, Leptosphaeria maculans, confers avirulence to Brassica juncea cultivars. Mol. Plant Pathol. 15, 523-530.
50. Van den Hooven, H.W., van den Burg, H.A., Vossen, P., Boeren, S., de Wit, P.J.G.M., and, Vervoot, J., (2001) Disulfide bound structure of the AVR9 elicitor of the fungal tomato pathogen Cladosporium fulvum: evidence for a cystine knot. Biochemistry, 40, 3458-3466.
51. Ve, T., Williams, S.J., Catanzariti, A.M., et al. (2013) Structures of the flax-rust effector AvrM reveal insights into the molecular basis of plant-cell entry and effector-triggered immunity. Proc. Natl Acad. Sci. USA, 110, 17594-17599.
52. Vinatzer, B.A., Teitzel, G.M., Lee, M.W., Jelenska, J., Hotton, S., Fairfax, K., Jenrette, J., and, Greenberg, J.T., (2006) The type III effector repertoire of Pseudomonas syringae pv. syringae B728a and its role in survival and disease on host and non-host plants. Mol. Microbiol. 62, 26-44.
53. Wang, C.I.A., Guncar, G., Forwood, J.K., et al. (2007) Crystal structures of flax rust avirulence proteins AvrL567-A and -D reveal details of the structural basis for flax disease resistance specificity. Plant Cell, 19, 2898-2912.
54. Wawra, S., Djamei, A., Albert, I., Nurnberger, T., Kahmann, R., and, van West, P., (2013) In vitro translocation experiments with RxLR-reporter fusion proteins of Avr1b from Phytophthora sojae and AVR3a from Phytophthora infestans fail to demonstrate specific autonomous uptake in plant and animal cells. Mol. Plant Microbe Interact. 26, 528-536.
55. Whisson, S.C., Boevink, P.C., Moleleki, L., et al. (2007) A translocation signal for delivery of oomycete effector proteins into host plant cells. Nature, 450, 115-119.
56. Xu, B., Munoz, I.G., Janson, J.C., and, Stahlberg, J., (2002) Crystallization and X-ray analysis of native and selenomethionyl β-mannanase Man5A from blue mussel, Mytilus edulis, expressed in Pichia pastoris. Acta Crystallogr. D Biol. Crystallogr. 58, 542-545.
57. Yaeno, T., Li, H., Chaparro-Garcia, A., Schornack, S., Koshiba, S., Watanabe, S., Kigawa, T., Kamoun, S., and, Shirasu, K., (2011) Phosphatidylinositol monophosphate-binding interface in the oomycete RXLR effector AVR3a is required for its stability in host cells to modulate plant immunity. Proc. Natl Acad. Sci. USA, 108, 14682-14687.
58. Zhang, Z.M., Zhang, X., Zhou, Z.R., Hu, H.Y., Liu, M., Zhou, B., and, Zhou, J., (2013) Solution structure of the Magnaporthe oryzae avirulence protein AvrPiz-t. J. Biomol. NMR, 55, 2019-2223.