Structural and functional studies of a phosphatidic acid-binding antifungal plant defensin MtDef4: Identification of an RGFRRR motif governing fungal cell entry

PLoS ONE

Volumn 8, Issue 12, 2013, Pages

  Sagaram, Uma Shankar ^a   El Mounadi, Kaoutar ^a   Buchko, Garry W ^b   Berg, Howard R ^a   Kaur, Jagdeep ^a   Pandurangi, Raghu S ^a   Smith, Thomas J ^a   Shah, Dilip M ^a  

^a DONALD DANFORTH PLANT SCIENCE CENTER   (United States)

^b PACIFIC NORTHWEST NATIONAL LABORATORY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

DEFENSIN; DEFENSIN 4; PHOSPHATIDIC ACID; UNCLASSIFIED DRUG;

AMINO ACID SEQUENCE; ARTICLE; BARREL MEDIC; CELL MEMBRANE PERMEABILITY; CONTROLLED STUDY; CYTOPLASM; FUNGAL CELL; FUNGAL MEMBRANE; FUSARIUM GRAMINEARUM; IN VITRO STUDY; INTERNALIZATION; NONHUMAN; PLANT FUNGUS INTERACTION; PROTEIN FUNCTION; PROTEIN LIPID INTERACTION; PROTEIN MOTIF; PROTEIN STRUCTURE;

AMINO ACID MOTIFS; AMINO ACID SEQUENCE; AMINO ACIDS; ANTIFUNGAL AGENTS; DEFENSINS; FUSARIUM; MEDICAGO TRUNCATULA; MODELS, MOLECULAR; MOLECULAR SEQUENCE DATA; PHOSPHATIDIC ACIDS; PLANT PROTEINS; PROTEIN BINDING; PROTEIN CONFORMATION; SEQUENCE ALIGNMENT; STATIC ELECTRICITY;

EID: 84891754133     PISSN: None     EISSN: 19326203     Source Type: Journal    
DOI: 10.1371/journal.pone.0082485     Document Type: Article

References (70)

1. Broekaert WF, Cammue BPA, De Bolle MFC, Thevissen K, De Samblanx GW et al. (1997) Antimicrobial peptides from plants. Critical Reviews in Plant Sciences 16: 297-323. doi:10.1080/713608148.
2. Thomma BPHJ, Cammue BPA, Thevissen K (2002) Plant defensins. Planta 216: 193-202. doi:10.1007/s00425-002-0902-6. PubMed: 12447532.
3. Lay FT, Anderson MA (2005) Defensins - components of the innate immune system in plants. Curr Protein Pept Sci 6: 85-101. doi: 10.2174/ 1389203053027575. PubMed: 15638771.
4. Carvalho Ade O, Gomes VM (2009) Plant defensins - prospects for the biological functions and biotechnological properties. Peptides 30: 1007-1020. doi:10.1016/j.peptides.2009.01.018. PubMed: 19428780.
5. Stotz HU, Thomson JG, Wang Y (2009) Plant defensins: defense, development and application. Plant Signal Behav 4: 1010-1012. doi: 10.4161/psb.4.11.9755. PubMed: 20009545.
6. Broekaert WF, Terras FR, Cammue BP, Osborn RW (1995) Plant defensins: novel antimicrobial peptides as components of the host defense system. Plant Physiol 108: 1353-1358. doi:10.1104/pp.108.4.1353. PubMed: 7659744.
7. Terras FR, Schoofs HM, De Bolle MF, Van Leuven F, Rees SB et al. (1992) Analysis of two novel classes of plant antifungal proteins from radish (Raphanus sativus L.) seeds. J Biol Chem 267: 15301-15309. PubMed: 1639777.
8. Kaur J, Sagaram US, Shah DM (2011) Can plant defensins be sused to engineer durable commercially useful resistance in crop plants? Fungal Biology Reviews 25: 128-135. doi:10.1016/j.fbr.2011.07.004.
9. Aerts AM, François IE, Cammue BP, Thevissen K (2008) The mode of antifungal action of plant, insect and human defensins. Cell Mol Life Sci 65: 2069-2079. doi:10.1007/s00018-008-8035-0. PubMed: 18360739.
10. Sagaram U, Kaur J, Shah D (2012) Antifungal plant defensins: structure-activity relationships and modes of action and biotech applications. In: K Rajasekaran . American Chemical Society. pp. 317-336.
11. Thevissen K, Cammue BP, Lemaire K, Winderickx J, Dickson RC et al. (2000) A gene encoding a sphingolipid biosynthesis enzyme determines the sensitivity of Saccharomyces cerevisiae to an antifungal plant defensin from dahlia (Dahlia merckii). Proc Natl Acad Sci U S A 97: 9531-9536. doi:10.1073/pnas.160077797. PubMed: 10931938.
12. Thevissen K, Ferket KK, François IE, Cammue BP (2003) Interactions of antifungal plant defensins with fungal membrane components. Peptides 24: 1705-1712. doi:10.1016/j.peptides.2003.09.014. PubMed: 15019201.
13. Thevissen K, Francois IE, Aerts AM, Cammue BP (2005) Fungal sphingolipids as targets for the development of selective antifungal therapeutics. Curr Drug Targets 6: 923-928. doi: 10.2174/138945005774912771. PubMed: 16375675.
14. Thevissen K, de Mello Tavares P, Xu D, Blankenship J, Vandenbosch D et al. (2012) The plant defensin RsAFP2 induces cell wall stress, septin mislocalization and accumulation of ceramides in Candida albicans. Mol Microbiol 84: 166-180. doi:10.1111/j.1365-2958.2012.08017.x. PubMed: 22384976.
15. Thevissen K, Osborn RW, Acland DP, Broekaert WF (2000) Specific binding sites for an antifungal plant defensin from dahlia (Dahlia merckii) on fungal cells are required for antifungal activity. Mol Plant Microbe Interact 13: 54-61. doi:10.1094/MPMI.2000.13.1.54. PubMed: 10656585.
16. Ramamoorthy V, Cahoon EB, Li J, Thokala M, Minto RE et al. (2008) Glucosylceramide synthase is essential for alfalfa defensin-mediated growth inhibition but not for pathogenicity of Fusarium graminearum. Molecular Microbiology 66: 771-786.
17. Lobo DS, Pereira IB, Fragel-Madeira L, Medeiros LN, Cabral LM et al. (2007) Antifungal Pisum sativum defensin 1 interacts with Neurospora crassa cyclin F related to the cell cycle. Biochemistry 46: 987-996. doi: 10.1021/bi061441j. PubMed: 17240982.
18. van der Weerden NL, Lay FT, Anderson MA (2008) The plant defensin, NaD1, enters the cytoplasm of Fusarium oxysporum hyphae. J Biol Chem 283: 14445-14452. doi:10.1074/jbc.M709867200. PubMed: 18339623.
19. van der Weerden NL, Hancock RE, Anderson MA (2010) Permeabilization of fungal hyphae by the plant defensin NaD1 occurs through a cell wall-dependent process. J Biol Chem 285: 37513-37520. doi:10.1074/jbc.M110.134882. PubMed: 20861017.
20. Yount NY, Yeaman MR (2004) Multidimensional signatures in antimicrobial peptides. Proc Natl Acad Sci U S A 101: 7363-7368. doi: 10.1073/pnas.0401567101. PubMed: 15118082.
21. De Samblanx GW, Goderis IJ, Thevissen K, Raemaekers R, Fant F et al. (1997) Mutational analysis of a plant defensin from radish (Raphanus sativus L.) reveals two adjacent sites important for antifungal activity. J Biol Chem 272: 1171-1179. doi:10.1074/jbc.272.2.1171. PubMed: 8995418.
22. Sagaram US, Pandurangi R, Kaur J, Smith TJ, Shah DM (2011) Structure-Activity determinants in antifungal plant defensins MsDef1 and MtDef4 with different modes of action against Fusarium graminearum. PLOS ONE 6: e18550. doi:10.1371/journal.pone.0018550. PubMed: 21533249.
23. Kaur J, Thokala M, Robert-Seilaniantz A, Zhao P, Peyret H et al. (2012) Subcellular targeting of an evolutionarily conserved plant defensin MtDef4.2 determines the outcome of plant-pathogen interaction in transgenic Arabidopsis. Mol Plant Pathol 13: 1032-1046. doi:10.1111/j.1364-3703.2012.00813.x. PubMed: 22776629.
24. Ramamoorthy V, Zhao X, Snyder AK, Xu J-R, Shah DM (2007) Two mitogen-activated protein kinase signalling cascades mediate basal resistance to antifungal plant defensins in Fusarium graminearum. Cell Microbiol 9: 1491-1506. doi:10.1111/j.1462-5822.2006.00887.x. PubMed: 17253976.
25. Almeida MS, Cabral KM, Kurtenbach E, Almeida FC, Valente AP (2002) Solution structure of Pisum sativum defensin 1 by high resolution NMR: plant defensins, identical backbone with different mechanisms of action. J Mol Biol 315: 749-757. doi:10.1006/jmbi. 2001.5252. PubMed: 11812144.
26. Almeida MS, Cabral KMS, Zingali RB, Kurtenbach E (2000) Characterization of two novel defensin peptides from pea (Pisum sativum) seeds. Arch Biochem Biophys 378: 278-286. doi:10.1006/ abbi.2000.1824. PubMed: 10860545.
27. Armougom F, Moretti S, Poirot O, Audic S, Dumas P et al. (2006) Expresso: automatic incorporation of structural information in multiple sequence alignments using 3D-Coffee. Nucleic Acids Res 34: W604-W608. doi:10.1093/nar/gkl092. PubMed: 16845081.
28. Honig B, Nicholls A (1995) Classical electrostatics in biology and chemistry. Science 268: 1144-1149. doi:10.1126/science.7761829. PubMed: 7761829.
29. Nicholls A, Honig B (1991) A rapid finite difference algorithm, utilizing successive over relaxation to solve the Poisson Boltzmann equation. J Comput Chem 12: 435-445. doi:10.1002/jcc.540120405.
30. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM et al. (2004) UCSF Chimera - A Visualization System for Exploratory Research and Analysis. J Comput Chem 25: 1605-1612. doi:10.1002/jcc.20084. PubMed: 15264254.
31. Lee B, Richards FM (1971) The interpretation of protein structures: estimation of static accesibility. J Mol Biol 55: 379-400. doi: 10.1016/0022-2836(71)90324-X. PubMed: 5551392.
32. Saff EB, Kuijlaars ABJ (1997) Distributing many points on a sphere. Mathematical Intelligencer 19: 5-11. doi:10.1007/BF03024423.
33. Bailey S (1994) The CCP4 suite: programs for protein crystallography. Acta Crystallogr D 50: 760-763. doi:10.1107/S0907444994003112. PubMed: 15299374.
34. Ellis JG, Dodds PN (2011) Showdown at the RXLR motif: Serious differences of opinion in how effector proteins from filamentous eukaryotic pathogens enter plant cells. Proc Natl Acad Sci U S A 108: 14381-14382. doi:10.1073/pnas. 1111668108. PubMed: 21856948.
35. Tyler BM (2009) Entering and breaking: virulence effector proteins of oomycete plant pathogens. Cell Microbiol 11: 13-20. doi:10.1111/j.1462-5822. 2008.01240.x. PubMed: 18783481.
36. Whisson SC, Boevink PC, Moleleki L, Avrova AO, Morales JG et al. (2007) A translocation signal for delivery of oomycete effector proteins into host plant cells. Nature 450: 115-118. doi:10.1038/nature06203. PubMed: 17914356.
37. Spelbrink RG, Dilmac N, Allen A, Smith TJ, Shah DM et al. (2004) Differential antifungal and calcium channel-blocking activity among structurally related plant defensins. Plant Physiol 135: 2055-2067. doi: 10.1104/pp.104.040873. PubMed: 15299136.
38. Kale SD, Gu B, Capelluto DG, Dou D, Feldman E et al. (2010) External lipid PI3P mediates entry of eukaryotic pathogen effectors into plant and animal host cells. Cell 142: 284-295. doi:10.1016/j.cell. 2010.06.008. PubMed: 20655469.
39. Yaeno T, Li H, Chaparro-Garcia A, Schornack S, Koshiba S et al. (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 U S A 108: 14682-14687. doi:10.1073/pnas. 1106002108. PubMed: 21821794.
40. Bloch C Jr., Patel SU, Baud F, Zvelebil MJ, Carr MD et al. (1998) 1H NMR structure of an antifungal γ-thionin protein SIα1: similarity to scorpion toxins. Proteins 32: 334-349. doi:10.1002/(SICI)1097-0134(19980815)32: 3. PubMed: 9715910.
41. Fant F, Vranken W, Broekaert W, Borremans F (1998) Determination of the three-dimensional solution structure of Raphanus sativus antifungal protein 1 by 1 H NMR. J Mol Biol 279: 257-270. doi:10.1006/jmbi.1998.1767. PubMed: 9636715.
42. Fant F, Vranken WF, Borremans FA (1999) The three-dimensional solution structure of Aesculus hippocastanum antimicrobial protein 1 determined by 1 H nuclear magnetic resonance. Proteins 37: 388-403. doi:10.1002/(SICI)1097- 0134(19991115)37:3. PubMed: 10591099.
43. Janssen BJ, Schirra HJ, Lay FT, Anderson MA, Craik DJ (2003) Structure of Petunia hybrida defensin 1, a novel plant defensin with five disulfide bonds. Biochemistry 42: 8214-8222. doi:10.1021/bi034379o. PubMed: 12846570.
44. Lay FT, Schirra HJ, Scanlon MJ, Anderson MA, Craik DJ (2003) The three-dimensional solution structure of NaD1, a new floral defensin from Nicotiana alata and its application to a homology model of the crop defense protein alfAFP. J Mol Biol 325: 175-188. doi:10.1016/S0022-2836(02)01103-8. PubMed: 12473460.
45. Lay FT, Mills GD, Poon IK, Cowieson NP, Kirby N et al. (2012) Dimerization of plant defensin NaD1 enhances its antifungal activity. J Biol Chem 287: 19961-19972. doi:10.1074/jbc.M111.331009. PubMed: 22511788.
46. Drin G, Déméné H, Temsamani J, Brasseur R (2001) Translocation of the pAntp peptide and its amphipathic analogue AP-2AL. Biochemistry 40: 1824-1834. doi:10.1021/bi002019k. PubMed: 11327845.
47. Frankel AD, Pabo CO (1988) Cellular uptake of the tat protein from human immunodeficiency virus. Cell 55: 1189-1193. doi: 10.1016/0092-8674(88)90263-2. PubMed: 2849510.
48. Green M, Loewenstein PM (1988) Autonomous functional domains of chemically synthesized human immunodeficiency virus tat transactivator protein. Cell 55: 1179-1188. doi: 10.1016/0092-8674(88)90262-0. PubMed: 2849509.
49. Kale SD, Tyler BM (2011) Entry of oomycete and fungal effectors into plant and animal host cells. Cell Microbiol 13: 1839-1848. doi:10.1111/j. 1462-5822.2011.01659.x. PubMed: 21819515.
50. Muñoz A, Marcos JF, Read ND (2012) Concentration-dependent mechanisms of cell penetration and killing by the de novo-designed antifungal hexapeptide PAF26. Molecular Microbiology 85: 89-106. doi: 10.1111/j.1365-2958. 2012.08091.x.
51. Richard JP, Melikov K, Vives E, Ramos C, Verbeure B et al. (2003) Cell-penetrating peptides. A reevaluation of the mechanism of cellular uptake. J Biol Chem 278: 585-590. PubMed: 12411431.
52. Alves ID, Jiao CY, Aubry S, Aussedat B, Burlina F et al. (2010) Cell biology meets biophysics to unveil the different mechanisms of penetratin internalization in cells. Biochim Biophys Acta 1798: 2231-2239. doi:10.1016/j.bbamem.2010.02.009. PubMed: 20152795.
53. Schmidt N, Mishra A, Lai GH, Wong GC (2010) Arginine-rich cell-penetrating peptides. FEBS Lett 584: 1806-1813. doi:10.1016/j.febslet. 2009.11.046. PubMed: 19925791.
54. Sun F, Kale SD, Azurmendi HF, Li D, Tyler BM et al. (2013) Structural basis for interactions of the Phytophthora sojae RxLR effector Avh5 with phosphatidylinositol 3-phosphate and for host cell entry. Mol Plant Microbe Interact 26: 330-344. PubMed: 23075041.
55. Athenstaedt K, Daum G (1999) Phosphatidic acid, a key intermediate in lipid metabolism. Eur J Biochem 266: 1-16. doi:10.1046/j.1432-1327.1999.00822.x. PubMed: 10542045.
56. Lemmon MA (2008) Membrane recognition by phospholipid-binding domains. Nat Rev Mol Cell Biol 9: 99-111. doi:10.1038/nrm2328. PubMed: 18216767.
57. Testerink C, Munnik T (2005) Phosphatidic acid: a multifunctional stress signaling lipid in plants. Trends Plant Sci 10: 368-375. doi: 10.1016/j.tplants.2005.06.002. PubMed: 16023886.
58. Wang X (2005) Regulatory functions of phospholipase D and phoshatidic acid in plant, growth, development and stress response. Plant Physiol 139: 566-573. doi:10.1104/pp.105.068809. PubMed: 16219918.
59. Dracatos PM, van der Weerden NL, Carroll KT, Johnson ED, Plummer KM et al. (2013) Inhibition of cereal rust fungi by both class I and II defensins derived from the flowers of Nicotiana alata. Mol Plant Pathol. doi:10.1111/mpp.12066.
60. Correll JC, Klittich CJR, Leslie JF (1987) Nitrate nonutilizing mutants of Fusarium graminearum and their use in vegetative compatibility tests. Phytopathology 77: 1640-1646. doi:10.1094/Phyto-77-1640.
61. Cappelini RA, Peterson JL (1965) Macroconidium formation in submerged cultures by a non-sporulating strain of Gibberella zeae. Mycologia 57: 962-966. doi:10.2307/3756895.
62. Broekaert WF, Terras FR, Cammue BP, Vanderleyden J (1990) An automated quantitative assay for fungal growth inhibition. FEMS Microbiology Letters 69: 55-60. doi:10.1111/j.1574-6968.1990.tb04174.x.
63. Rodriguez E, Krishna NR (2001) An economical method for (15)N/ (13)C isotopic labeling of proteins expressed in Pichia pastoris. J Biochem 130: 19-22. doi:10.1093/oxfordjournals.jbchem.a002957. PubMed: 11432775.
64. Goddard TD, Kneller DG (2008) SPARKY 3. San Francisco: University of California.
65. Wishart DS, Bigam CG, Yao J, Abildgaard F, Dyson HJ et al. (1995) 1H, 13C and 15N Chemical shift referencing in biomolecular NMR. J Biomol NMR 6: 135-140. PubMed: 8589602.
66. Güntert P (2004.) Automated NMR structure calculation with CYANA. Methods Mol Biol 278: 353-378. PubMed: 15318003.
67. Cornilescu G, Delaglio F, Bax A (1999) Protein backbone angle restraints from searching a database for chemical shift and sequence homology. J Biomol NMR 13: 289-301. doi:10.1023/A:1008392405740. PubMed: 10212987.
68. Linge JP, Nilges M (1999) Influence of non-bonded parameters on the quality of NMR structures: A new force field for NMR structure calculation. J Biomol NMR 13: 51-59. doi:10.1023/A:1008365802830. PubMed: 10905826.
69. Bhattacharya A, Tejero R, Montelione GT (2007) Evaluating protein structures determined by structural genomics consortia. Proteins 66: 778-795. PubMed: 17186527.
70. Guo L, Mishra G, Taylor K, Wang X (2011) Phosphatidic acid binds and stimulates Arabidopsis sphingosine kinases. J Biol Chem 286: 13336-13345. doi:10.1074/jbc.M110.190892. PubMed: 21330371.