Vesicular trafficking plays a crucial role in protein localization and movement, signal transduction, and multiple developmental processes in eukaryotic cells. Vesicle fusion is the final and key step in vesicle-mediated trafficking and mainly relies on SNAREs (soluble N-ethylmaleimide-sensitive factor attachment protein receptors), the regulators including SM (Sec1/Munc18) family proteins, Rab GTPases and exocyst subunits. Verticillium dahliae is a widespread soil fungus that causes disruptive vascular diseases on a wide range of plants. To date, no genes involved in vesicular fusion process have been identified and characterized in V. dahliae. The recent publication of the draft genome sequence of V. dahliae allowed us to conduct a genome-wide identification, phylogeny and expression profile of genes encoding vesicular fusion components. Using compared genomics and phylogenetic methods, we identified 44 genes encoding vesicle fusion components in the V. dahliae genome. According to the structural features of their encoded proteins, the 44 V. dahliae genes were classified into 22 SNAREs (6 Qa-, 4 Qb-, 6 Qc-, 1 Qbc- and 5 R-types), 4 SM family proteins, 10 Rab GTPases and 8 exocyst proteins. Based on phylogeny and motif constitution analysis, orthologs of vesicle fusion component in filamentous fungi were generally clustered together into the same subclasses with well-supported bootstrap values. Analysis of the expression profiles of these genes indicated that many of them are significantly differentially expressed during vegetative growth and microsclerotia formation in V. dahliae. The analysis show that many components of vesicle fusion are well conserved in filamentous fungi and indicate that vesicle fusion plays a critical role in microsclerotia formation of smoke tree wilt fungus V. dahliae. The genome-wide identification and expression analysis of components involved in vesicle fusion should facilitate research in this gene family and give new insights toward elucidating their functions in growth, development and pathogenesis of V. dahliae.
References
[1]
Yao HY, Xue HW (2011) Signals and mechanisms affecting vesicular trafficking during root growth. Curr Opin Plant Biol 14: 571–579.
[2]
Bonifacino JS, Glick BS (2004) The mechanisms of vesicle budding and fusion. Cell 116: 153–166.
[3]
Samaj J, Muller J, Beck M, Bohm N, Menzel D (2006) Vesicular trafficking, cytoskeleton and signalling in root hairs and pollen tubes. Trends Plant Sci 11: 594–600.
[4]
Wickner W (2010) Membrane fusion: five lipids, four SNAREs, three chaperones, two nucleotides, and a Rab, all dancing in a ring on yeast vacuoles. Annu Rev Cell Dev Biol 26: 115–136.
[5]
Gerst JE (1999) SNAREs and SNARE regulators in membrane fusion and exocytosis. Cell Mol Life Sci 55: 707–734.
[6]
Ostrowicz CW, Meiringer CT, Ungermann C (2008) Yeast vacuole fusion: a model system for eukaryotic endomembrane dynamics. Autophagy 4: 5–19.
[7]
Koumandou VL, Dacks JB, Coulson RM, Field MC (2007) Control systems for membrane fusion in the ancestral eukaryote; evolution of tethering complexes and SM proteins. BMC Evol Biol 7: 29.
[8]
Lipka V, Kwon C, Panstruga R (2007) SNARE-ware: the role of SNARE-domain proteins in plant biology. Annu Rev Cell Dev Biol 23: 147–174.
[9]
Li L, Chin LS (2003) The molecular machinery of synaptic vesicle exocytosis. Cell Mol Life Sci 60: 942–960.
[10]
Ungar D, Hughson FM (2003) SNARE protein structure and function. Annu Rev Cell Dev Biol 19: 493–517.
Ferro-Novick S, Jahn R (1994) Vesicle fusion from yeast to man. Nature 370: 191–193.
[13]
Kloepper TH, Kienle CN, Fasshauer D (2007) An elaborate classification of SNARE proteins sheds light on the conservation of the eukaryotic endomembrane system. Mol Biol Cell 18: 3463–3471.
Fasshauer D, Sutton RB, Brunger AT, Jahn R (1998) Conserved structural features of the synaptic fusion complex: SNARE proteins reclassified as Q- and R-SNAREs. Proc Natl Acad Sci U S A 95: 15781–15786.
[16]
Burri L, Lithgow T (2004) A complete set of SNAREs in yeast. Traffic 5: 45–52.
[17]
Kienle N, Kloepper TH, Fasshauer D (2009) Phylogeny of the SNARE vesicle fusion machinery yields insights into the conservation of the secretory pathway in fungi. BMC Evol Biol 9: 19.
[18]
Ayong L, Pagnotti G, Tobon AB, Chakrabarti D (2007) Identification of Plasmodium falciparum family of SNAREs. Mol Biochem Parasitol 152: 113–122.
[19]
Sanderfoot A (2007) Increases in the number of SNARE genes parallels the rise of multicellularity among the green plants. Plant Physiol 144: 6–17.
[20]
Bock JB, Matern HT, Peden AA, Scheller RH (2001) A genomic perspective on membrane compartment organization. Nature 409: 839–841.
[21]
Hong W (2005) SNAREs and traffic. Biochim Biophys Acta 1744: 120–144.
[22]
Kuratsu M, Taura A, Shoji JY, Kikuchi S, Arioka M, et al. (2007) Systematic analysis of SNARE localization in the filamentous fungus Aspergillus oryzae. Fungal Genet Biol 44: 1310–1323.
[23]
Morgera F, Sallah MR, Dubuke ML, Gandhi P, Brewer DN, et al. (2012) Regulation of exocytosis by the exocyst subunit Sec6 and the SM protein Sec1. Mol Biol Cell 23: 337–346.
[24]
Gupta GD, Brent Heath I (2002) Predicting the distribution, conservation, and functions of SNAREs and related proteins in fungi. Fungal Genet Biol 36: 1–21.
[25]
Novick P, Zerial M (1997) The diversity of Rab proteins in vesicle transport. Curr Opin Cell Biol 9: 496–504.
[26]
Pereira-Leal JB, Seabra MC (2001) Evolution of the Rab family of small GTP-binding proteins. J Mol Biol 313: 889–901.
[27]
Pereira-Leal JB (2008) The Ypt/Rab family and the evolution of trafficking in fungi. Traffic 9: 27–38.
[28]
Elias M, Brighouse A, Gabernet-Castello C, Field MC, Dacks JB (2012) Sculpting the endomembrane system in deep time: high resolution phylogenetics of Rab GTPases. J Cell Sci 125: 2500–2508.
[29]
Bassham DC, Brandizzi F, Otegui MS, Sanderfoot AA (2008) The secretory system of Arabidopsis. Arabidopsis Book 6: e0116.
[30]
Toonen RF, Verhage M (2003) Vesicle trafficking: pleasure and pain from SM genes. Trends Cell Biol 13: 177–186.
[31]
Misura KM, Scheller RH, Weis WI (2000) Three-dimensional structure of the neuronal-Sec1-syntaxin 1a complex. Nature 404: 355–362.
[32]
Shandala T, Kakavanos-Plew R, Ng YS, Bader C, Sorvina A, et al.. (2012) Molecular Machinery Regulating Exocytosis, Crosstalk and Integration of Membrane Trafficking Pathways, Dr. Roberto Weigert (Ed.), ISBN: 978–953–51–0515–2, InTech, DOI: 10.5772/39110.
[33]
TerBush DR, Maurice T, Roth D, Novick P (1996) The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae. EMBO J 15: 6483–6494.
[34]
Guo W, Tamanoi F, Novick P (2001) Spatial regulation of the exocyst complex by Rho1 GTPase. Nat Cell Biol 3: 353–360.
[35]
McNew JA (2008) Regulation of SNARE-mediated membrane fusion during exocytosis. Chem Rev 108: 1669–1686.
Klosterman SJ, Atallah ZK, Vallad GE, Subbarao KV (2009) Diversity, Pathogenicity, and Management of Verticillium Species. Annu Rev Phytopathol 47: 39–62.
[38]
Fradin EF, Thomma BP (2006) Physiology and molecular aspects of Verticillium wilt diseases caused by V. dahliae and V. albo-atrum. Mol Plant Pathol 7: 71–86.
Friebertshauser GE, DeVay JE (1982) Differential Effects of the Defoliating and Nondefoliating Pathotypes of Verticillium dahliae Upon the Growth and Development of Gossypium hirsutum. Phytopathology 72: 872–877.
[41]
Koike ST, Subbarao KV, Davis RM, R.Gordon T, C.Hubbard J (1994) Verticillium Wilt of Cauliflower in California. Plant Dis 78: 1116–1121.
[42]
Johnson D, Dung J (2010) Verticillium wilt of potato - the pathogen, disease and management. Can J Plant Pathol 32: 58–67.
[43]
Wang J, Tian C, Ge J (2008) Research progress of the tree wilt disease caused by Verticillium. Forest Pest and Disease 10: 30–36 (in Chinese)..
[44]
Hou YY, Shi FM, Li YZ (2008) Verticillium dahliae toxins induce changes in expression of a putative histone H2B and a pumilio/Puf RNA-binding protein in Arabidopsis thaliana. J Plant Pathol 90: 117–120.
[45]
Palmer CS, Saleeba JA, Lyon BR (2005) Phytotoxicity on cotton ex-plants of an 18.5 kDa protein from culture filtrates of Verticillium dahliae. Physiol Mol Plant Pathol 67: 308–318.
[46]
Wang JY, Cai Y, Gou JY, Mao YB, Xu YH, et al. (2004) VdNEP, an elicitor from Verticillium dahliae, induces cotton plant wilting. Appl Environ Microbiol 70: 4989–4995.
[47]
Zhou BJ, Jia PS, Gao F, Guo HS (2012) Molecular characterization and functional analysis of a necrosis- and ethylene-inducing, protein-encoding gene family from Verticillium dahliae. Mol Plant Microbe Interact 25: 964–975.
[48]
Klimes A, Dobinson KF (2006) A hydrophobin gene, VDH1, is involved in microsclerotial development and spore viability in the plant pathogen Verticillium dahliae. Fungal Genet Biol 43: 283–294.
[49]
Perry JW, Evert RF (1982) Structure of microsclerotia of Verticillium dahliae in roots of ‘Russett Burbank’ potatoes. Can J Bot 62: 396–401.
[50]
Griffiths DA (1970) The fine structure of developing microsclerotia of Verticillium dahliae Kleb. Arch Mikrobiol 74: 207–212.
[51]
Song W, Dou X, Qi Z, Wang Q, Zhang X, et al. (2010) R-SNARE homolog MoSec22 is required for conidiogenesis, cell wall integrity, and pathogenesis of Magnaporthe oryzae. PLoS One 5: e13193.
[52]
Dou X, Wang Q, Qi Z, Song W, Wang W, et al. (2011) MoVam7, a conserved SNARE involved in vacuole assembly, is required for growth, endocytosis, ROS accumulation, and pathogenesis of Magnaporthe oryzae. PLoS One 6: e16439.
[53]
Klosterman SJ, Subbarao KV, Kang S, Veronese P, Gold SE, et al. (2011) Comparative Genomics Yields Insights into Niche Adaptation of Plant Vascular Wilt Pathogens. PLoS Pathog 7: e1002137.
[54]
Dobinson KF, Lecomte N, Lazarovits G (1997) Production of an extracellular trypsin-like protease by the fungal plant pathogen Verticillium dahliae. Can J Microbiol 43: 227–233.
[55]
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, et al. (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23: 2947–2948.
[56]
Guindon S, Dufayard JF, Lefort V, Anisimova M, Hordijk W, et al. (2010) New algorithms and methods to estimate maximum-likelihood phylogenies: assessing the performance of PhyML 3.0. Syst Biol 59: 307–321.
[57]
Wang Y, Xiao S, Xiong D, Tian C (2013) Genetic transformation, infection process and qPCR quantification of Verticillium dahliae on smoke-tree Cotinus coggygria. Australasian Plant Pathol 42: 33–41.
[58]
Tzima AK, Paplomatas EJ, Tsitsigiannis DI, Kang S (2012) The G protein beta subunit controls virulence and multiple growth- and development-related traits in Verticillium dahliae. Fungal Genet Biol 49: 271–283.
[59]
Klimes A, Amyotte SG, Grant S, Kang S, Dobinson KF (2008) Microsclerotia development in Verticillium dahliae: Regulation and differential expression of the hydrophobin gene VDH1. Fungal Genet Biol 45: 1525–1532.
[60]
Agrawal GK, Jwa NS, Lebrun MH, Job D, Rakwal R (2010) Plant secretome: unlocking secrets of the secreted proteins. Proteomics 10: 799–827.
[61]
Girard V, Dieryckx C, Job C, Job D (2013) Secretomes: the fungal strike force. Proteomics 13: 597–608.
[62]
de Jonge R, Bolton MD, Thomma BP (2011) How filamentous pathogens co-opt plants: the ins and outs of fungal effectors. Curr Opin Plant Biol 14: 400–406.
[63]
Oliva R, Win J, Raffaele S, Boutemy L, Bozkurt TO, et al. (2010) Recent developments in effector biology of filamentous plant pathogens. Cell Microbiol 12: 705–715.
[64]
Wickner W (2002) Yeast vacuoles and membrane fusion pathways. EMBO J 21: 1241–1247.
[65]
Wedlich-Soldner R, Bolker M, Kahmann R, Steinberg G (2000) A putative endosomal t-SNARE links exo- and endocytosis in the phytopathogenic fungus Ustilago maydis. EMBO J 19: 1974–1986.
[66]
Rauyaree P, Ospina-Giraldo MD, Kang S, Bhat RG, Subbarao KV, et al. (2005) Mutations in VMK1, a mitogen-activated protein kinase gene, affect microsclerotia formation and pathogenicity in Verticillium dahliae. Curr Genet 48: 109–116.
[67]
Gao F, Zhou B, Li G, Jia P, Li H, et al. (2010) A Glutamic Acid-Rich Protein Identified in Verticillium dahliae from an Insertional Mutagenesis Affects Microsclerotial Formation and Pathogenicity. PLoS ONE 5: e15319.