Skip to main content
SpringerLink
Log in

Molecular cloning and characterization of cDNA encoding a Translocon-Associated Protein (TRAPδ) from the root-lesion nematode Pratylenchus goodeyi

  • Published:
European Journal of Plant Pathology Aims and scope Submit manuscript

Abstract

The translocon-associated protein (TRAP) complex comprises four subunits (α, β, γ, δ) and is located in the endoplasmic reticulum membrane at translocation sites. The TRAP complex is required for the efficient translocation of substrates and to correct or eliminate misfolded proteins. In this study, we described the cloning and characterization of a cDNA encoding a TRAP from the phytoparasitic nematode Pratylenchus goodeyi (Pg). The full-length cDNA had an estimated size of 690 bp and encodes a 177 amino acid peptide. The deduced protein after sequence analysis codes for TRAPδ subunit homologous to TRAPδ from other nematodes. The Pg-TRAPδ had a signal peptide indicating a possible involvement in the transport and binding of other proteins at the endoplasmic reticulum membrane. The increase in relative expression of Pg-trapδ, assessed by semi-quantitative PCR, was induced over time in nematodes exposed to a nematostatic/nematicide extract of Solanum nigrum, suggesting that this gene product might be influenced by response mechanisms to stress in P. goodeyi. This is the first report of the cloning and characterization of trap cDNA from plant endoparasitic nematodes.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Abràmoff, M. D., Magalhães, P. J., & Ram, S. J. (2004). Image Processing with Image J. Biophotonics International, 11, 36–42.

    Google Scholar 

  • Abrantes, I. M., de, O., de Morais, M. M. N., Paiva, I. M. P., de, F. R., Santos, M. S. N., & de, A. (1976). Análise nematológica de solos e plantas. Ciência Biológica (Portugal), 1, 139–155.

    Google Scholar 

  • Altschul, S. F., Madden, T. L., Schäffer, A. A., Zhang, J., Zhang, Z., Miller, W., et al. (1997). Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Research, 25, 3389–3402.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Atkinson, H. J., Urwin, P. E., & McPherson, M. J. (2003). Engineering plants for nematode resistance. Annual Review of Phytopathology, 41, 615–639.

    Article  CAS  PubMed  Google Scholar 

  • Bendtsen, J. D., Nielsen, H., von Heijne, G., & Brunak, S. (2004). Improved prediction of signal peptides: SignalP 3.0. Journal of Molecular Biology, 340, 783–795.

    Article  PubMed  Google Scholar 

  • Benson, D. A., Cavanaugh, M., Clark, K., Karsch-Mizrachi, I., Lipman, D. J., Ostell, J., et al. (2013). GenBank. Nucleic Acids Research, 41, D36–D42.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • da Rocha Fragoso, R., Lourenço, I. T., Batista, J. A. N., Oliveira-Neto, O. B., Silva, M. C. M., Rocha, T. L., et al. (2009). Meloidogyne incognita: Molecular cloning and characterization of cDNA encoding a cathepsin D-like aspartic proteinase. Experimental Parasitology, 121, 115–123.

    Article  CAS  PubMed  Google Scholar 

  • Davis, E. L., Hussey, R. S., Baum, T. J., Bakker, J., Schots, A., Rosso, M. N., et al. (2000). Nematode parasitism genes. Annual Review of Phytopathology, 38, 365–396.

    Article  CAS  PubMed  Google Scholar 

  • De Luca, F., de Giorgi, C., Di Vito, M., & Lamberti, F. (1996). Sequence analysis of cut-1 gene and of its flanking regions in Meloidogyne artiellia. Nematologia Mediterranea, 24, 125–128.

    Google Scholar 

  • De Luca, F., Reyes, A., Troccoli, A., & Castillo, P. (2011). Molecular variability and phylogenetic relationships among different species and populations of Pratylenchus (Nematoda: Pratylenchidae) as inferred from the analysis of the ITS rDNA. European Journal of Plant Pathology, 130, 415–426.

    Article  Google Scholar 

  • Fons, R. D., Bogert, B. A., & Hegde, R. S. (2003). Substrate-specific function of the translocon-associated protein complex during translocation across the ER membrane. The Journal of Cell Biology, 160, 529–539.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Gasteiger, E., Hoogland, C., Gattiker, A., Duvaud, S., Wilkins, M. R., Appel, R. D., et al. (2005). Protein identification and analysis tools on the ExPASy server. In: Walker, J. M., (Ed.), The proteomics protocols handbook. Humana Press. pp. 571–607

  • Gowen, S., & Quénéhervé, P. (1990). Nematode parasites of bananas, plantains and abaca. In M. Luc, R. A. Sikora, & J. Bridge (Eds.), Plant parasitic nematodes in subtropical and tropical agriculture (pp. 431–460). Wallingford: CAB International.

    Google Scholar 

  • Guda, C. (2006). pTARGET: A web server for predicting protein subcellular localization. Nucleic Acids Research, 35, 210–213.

    Article  Google Scholar 

  • Guda, C., & Subramaniam, S. (2005). pTARGET: A new method for predicting protein subcellular localization in eukaryotes. Bioinformatics, 21, 3963–3969.

    Article  CAS  PubMed  Google Scholar 

  • Hajarnavis, A., & Durbin, R. (2006). A conserved sequence motif in 3′ untranslated regions of ribosomal protein mRNAs in nematodes. RNA, 12, 1786–1789.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hartmann, E., Görlich, D., Kostka, S., Otto, A., Kraft, R., Knespel, S., et al. (1993). A tetrameric complex of membrane proteins in the endoplasmic reticulum. European Journal of Biochemistry, 214, 375–381.

    Article  CAS  PubMed  Google Scholar 

  • Hegde, R. S., & Kang, S. W. (2008). The concept of translocational regulation. The Journal of Cell Biology, 182, 225–232.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Hiss, J. A., & Schneider, G. (2009). Architecture, function and prediction of long signal peptides. Briefings in Bioinformatics, 10, 569–578.

    Article  CAS  PubMed  Google Scholar 

  • Hooper, D. J. (1986). Extraction of nematodes from plant material. In J. F. Southey (Ed.), Laboratory methods for work with plant and soil nematodes (pp. 51–58). London: Her Majesty’s Stationery Office.

    Google Scholar 

  • Hulbert, S. H., Webb, C. A., Smith, S. M., & Sun, Q. (2001). Resistance gene complexes: evolution and utilization. Annual Review of Phytopathology, 39, 285–312.

    Article  CAS  PubMed  Google Scholar 

  • Jensen, L. J., Skovgaard, M., & Brunak, S. (2002). Prediction of novel archaeal enzymes from sequence-derived features. Protein Science, 11, 2894–2898.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Käll, L., Krogh, A., & Sonnhammer, E. L. L. (2007). Advantages of combined transmembrane topology and signal peptide prediction-the Phobius web server. Nucleic Acids Research, 35, 429–432.

    Article  Google Scholar 

  • Kapp, K., Schrempf, S., Lemberg, M. K. & Dobberstein, B. (2000). Post-targeting functions of signal peptides. In: Madame Curie Bioscience Database [Internet]. Austin (TX): Landes Bioscience. Available from: http://www.ncbi.nlm.nih.gov/books/NBK6322/. [accessed 22 April].

  • Kozak, M. (1989). The scanning model for translation: an update. The Journal of Cell Biology, 108, 229–241.

    Article  CAS  PubMed  Google Scholar 

  • Laplaze, L., Ribeiro, A., Franche, C., Duhoux, E., Auguy, F., Bogusz, D., et al. (2000). Characterization of a Casuarina glauca nodule-specific subtilisin-like protease gene, a homolog of Alnus glutinosa ag12. Molecular Plant-Microbe Interactions, 13, 113–117.

    Article  CAS  PubMed  Google Scholar 

  • Launikonis, B. S., & Stephenson, D. G. (1997). Effect of saponin treatment on the sarcoplasmic reticulum of rat, cane toad and crustacean (Yabby) skeletal muscle. The Journal of Physiology, 504, 425–437.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Martínez-Gil, L., Saurí, A., Martin-Renom, M. A., & Mingarro, I. (2011). Membrane protein integration into the endoplasmatic reticulum. FEBS Journal, 278, 3846–3858.

    Article  PubMed  Google Scholar 

  • Meldal, B. H. M., Debenham, N. J., De Ley, P., De Ley, I. T., Vanfleteren, J. R., Vierstraete, A. R., et al. (2007). An improved molecular phylogeny of the Nematoda with special emphasis on marine taxa. Molecular Phylogenetics and Evolution, 42, 622–636.

    Article  CAS  PubMed  Google Scholar 

  • Ménétret, J. F., Hegde, R. S., Heinrich, S. U., Chandramouli, P., Ludtke, S. J., Rapoport, T. A., et al. (2005). Architecture of the ribosome–channel complex derived from native membranes. Journal of Molecular Biology, 348, 445–457.

    Article  PubMed  Google Scholar 

  • Ménétret, J. F., Hegde, R. S., Aguiar, M., Gygi, S. P., Park, E., Rapoport, T. A., et al. (2008). Single copies of Sec61 and TRAP associate with a non-translating mammalian ribosome. Structure, 16, 1126–1137.

    Article  PubMed Central  PubMed  Google Scholar 

  • Mesbah, K., Camus, A., Babinet, C., & Barra, J. (2006). Mutation in the Trapα/Ssr1 gene, encoding translocon-associated protein. Results in outflow tract morphogenetic defects. Molecular Cell Biology, 26, 7760–7771.

    Article  CAS  Google Scholar 

  • Miyazaki, K., Fujita, T., Ozaki, T., Kato, C., Kurose, Y., Sakamoto, M., et al. (2004). NEDL1, a novel ubiquitin-protein isopeptide ligase for dishevelled-1, targets mutant superoxide dismutase-1. Journal of Biological Chemistry, 297, 11327–11335.

    Article  Google Scholar 

  • Nagasawa, K., Higashi, T., Hosokawa, N., Kaufman, R. J., & Nagata, K. (2007). Simultaneous induction of the four subunits of the TRAP complex by ER stress accelerates ER degradation. EMBO reports, 8, 483–489.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Nakai, K., & Kanehisa, M. (1992). A knowledge base for predicting protein localization sites in eukaryotic cells. Genomics, 14, 897–911.

    Article  CAS  PubMed  Google Scholar 

  • Pestana, M., Rodrigues, M., Teixeira, L., de O Abrantes, I. M., Gouveia, M., & Cordeiro, N. (2013). In vitro evaluation of nematicidal properties of Solanum sisymbriifolium and S nigrum extracts on Pratylenchus goodeyi. Nematology. doi:10.1163/15685411-00002743.

    Google Scholar 

  • Pinochet, J., Fernandez, C., & Sarah, J. L. (1995). Influence of temperature on in vitro reproduction of Pratylenchus coffeae, P. goodeyi and Radopholus similis. Fundamental and Applied Nematology, 18, 391–392.

    Google Scholar 

  • Quevillon, E., Silventoinen, V., Pillai, S., Harte, N., Mulder, N., Apweiler, R., et al. (2005). InterProScan: protein domains identifier. Nucleic Acids Research, 33, W116–W120.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Rosso, M. N., Favery, B., Piotte, C., Arthaud, L., De Boer, J. M., Hussey, R. S., et al. (1999). Isolation of a cDNA encoding a β-1,4-endoglucanase in the root-knot nematode Meloidogyne incognita and expression analysis during plant parasitism. Molecular Plant-Microbe Interactions, 12, 585–591.

    Article  CAS  PubMed  Google Scholar 

  • Rost, B., & Liu, J. (2003). The PredictProtein server. Nucleic Acids Res., 31, 3300–3304.

    Article  CAS  Google Scholar 

  • Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual (2nd ed.). New York: Cold Spring Harbor Laboratory Press.

    Google Scholar 

  • Sher, S. A., & Allen, M. W. (1953). Revision of the genus Pratylenchus (Nematoda: Tylenchidae). University of California Publications in Zoology, 57, 441–469.

    Google Scholar 

  • Stover, R., & Simmonds, N. W. (1987). Bananas. Essex: Longman scientific and technical, harlow. 468 pp.

    Google Scholar 

  • Thomas-Hall, S., Campbell, P. R., Carlens, K., Kawanish, E., Swennen, R., Sági, L., et al. (2007). Phylogenetic and molecular analysis of the ribulose-1,5-bisphosphate carboxylase small subunit gene family in banana. Journal of Experimental Botany, 58, 2685–2697.

    Article  CAS  PubMed  Google Scholar 

  • Waeyenberge, L., Ryss, A., Moens, M., Pinochet, J., & Vrain, T. C. (2000). Molecular characterization of 18 Pratylenchus species using rDNA restriction fragment length polymorphism. Nematology, 2, 135–142.

    Article  CAS  Google Scholar 

  • Wassler, M., Jonasson, I., Persson, R., & Fries, E. (1987). Differential permeabilization of membranes by saponin treatment of isolated rat hepatocytes. Biochemical Journal, 247, 407–415.

    CAS  PubMed Central  PubMed  Google Scholar 

Download references

Acknowledgments

M. Pestana thanks to CITMA for the awarding of a Doctoral grant (Project n.º 001080/2010/132).

Author information

Authors and Affiliations

  1. Madeira Agricultural Laboratory, RAM, 9135-372, Santa Cruz, Portugal

    Margarida Pestana

  2. Centre of Life Sciences, University of Madeira, 9000-390, Funchal, Portugal

    Margarida Pestana & Manuela Gouveia

  3. IMAR-CMA, Department of Life Sciences, University of Coimbra, 3004-517, Coimbra, Portugal

    Isabel M. de O. Abrantes

Authors
  1. Margarida Pestana
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Isabel M. de O. Abrantes
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Manuela Gouveia
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Margarida Pestana.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pestana, M., de O. Abrantes, I.M. & Gouveia, M. Molecular cloning and characterization of cDNA encoding a Translocon-Associated Protein (TRAPδ) from the root-lesion nematode Pratylenchus goodeyi . Eur J Plant Pathol 139, 289–298 (2014). https://doi.org/10.1007/s10658-013-0363-8

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10658-013-0363-8

Keywords

Search

Navigation