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The impact on microtubule network of a bracovirus IκB-like protein

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Abstract

Polydnavirus-encoded IκB-like proteins are similar to insect and mammalian IκB, and an immunosuppressive function in the host cells has been inferred to these proteins. Here we show that the expression of one of these IκB-like viral genes, the TnBVank1, in the Drosophila germline affects the localization of gurken, bicoid, and oskar mRNAs whose gene products are relevant for proper embryonic patterning. The altered localization of these mRNAs is suggestive of general defects in the intracellular, microtubule-based, trafficking routes. Analysis of microtubule motor proteins components such as the dynein heavy chain and the kinesin heavy chain revealed defects in the polarized microtubule network. Interestingly, the TnBVANK1 viral protein is uniformly distributed over the entire oocyte cortex, and appears to be anchored to the microtubule ends. Our data open up a very interesting issue on novel function(s) played by the ank gene family by interfering with cytoskeleton organization.

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References

  1. Pennacchio F, Strand MR (2006) Evolution of developmental strategies in parasitic hymenoptera. Annu Rev Entomol 51:233–258

    Article  CAS  PubMed  Google Scholar 

  2. Webb BA, Strand MR (2005) The biology and genomics of polydnaviruses. In: Gilbert LI, Iatrou K, Gill SS (eds) Comprehensive molecular insect science, vol 6. Elsevier, San Diego, pp 323–360

  3. Webb BA, Beckage NE, Hayakawa Y, Krell PJ, Lanzrein B, Stoltz DB, Strand MR, Summers MD (2000) Family Polydnaviridae. In: van Regenmortel MHV, Fauquet CM, Bishop DHL, Carstens EB, Estes MK, Lemon SM, Maniloff J, Mayo MA, McGeoch DJ, Pringle CR, Wickner RB et al (eds) Virus taxonomy: seventh report of the international committee on taxonomy of viruses. Academic Press, San Diego, pp 253–260

    Google Scholar 

  4. Deng L, Stoltz DB, Webb BA (2000) A gene encoding a polydnavirus structural polypeptide is not encapsidated. Virology 269:440–450

    Article  CAS  PubMed  Google Scholar 

  5. Bézier A, Annaheim M, Herbinière J, Wetterwald C, Gyapay G, Bernard-Samain S, Wincker P, Roditi I, Heller M, Belghazi M, Pfister-Wilhem R, Periquet G, Dupuy C, Huguet E, Volkoff AN, Lanzrein B, Drezen JM (2009) Polydnaviruses of braconid wasps derive from an ancestral nudivirus. Science 323:926–930

    Article  PubMed  Google Scholar 

  6. Bézier A, Herbinière J, Lanzrein B, Drezen JM (2009) Polydnavirus hidden face: the genes producing virus particles of parasitic wasps. J Invertebr Pathol 101:194–203

    Article  PubMed  Google Scholar 

  7. Espagne E, Dupuy C, Huguet E, Cattolico L, Provost B, Martins N, Poirié M, Periquet G, Drezen JM (2004) Genome sequence of a polydnavirus: insights into symbiotic virus evolution. Science 306:286–289

    Article  CAS  PubMed  Google Scholar 

  8. Silverman N, Maniatis T (2001) NF-κB signaling pathways in mammalian and insect innate immunity. Genes Dev 15:2321–2342

    Article  CAS  PubMed  Google Scholar 

  9. Kroemer JA, Webb BA (2005) IκB-related vankyrin genes in the Campoletis sonorensis ichnovirus: temporal and tissue-specific patterns of expression in parasitized Heliothis virescens lepidopteran hosts. J Virol 79:7617–7628

    Article  CAS  PubMed  Google Scholar 

  10. Thoetkiattikul H, Beck MH, Strand MR (2005) Inhibitor κB-like proteins from a polydnavirus inhibit NF-κB activation and suppress the insect immune response. Proc Natl Acad Sci USA 102:11426–11431

    Article  CAS  PubMed  Google Scholar 

  11. Falabella P, Varricchio P, Provost B, Espagne E, Ferrarese R, Grimaldi A, de Eguileor M, Fimiani G, Ursini MV, Malva C, Drezen JM, Pennacchio F (2007) Characterization of the IκB-like gene family in polydnaviruses associated with wasps belonging to different Braconid subfamilies. J Gen Virol 88:92–104

    Article  CAS  PubMed  Google Scholar 

  12. Lapointe R, Tanaka K, Barney WE, Whitfield JB, Banks JC, Béliveau C, Stoltz D, Webb BA, Cusson M (2007) Genomic and morphological features of a banchine polydnavirus: comparison with bracoviruses and ichnoviruses. J Virol 81:6491–6501

    Article  CAS  PubMed  Google Scholar 

  13. Tian SP, Zhang JH, Wang CZ (2007) Cloning and characterization of two Campoletis chlorideae ichnovirus vankyrin genes expressed in parasitized host Helicoverpa armigera. J Insect Physiol 53:699–707

    Article  CAS  PubMed  Google Scholar 

  14. Shi M, Chen YF, Huang F, Liu PC, Zhou XP, Chen XX (2008) Characterization of a novel gene encoding ankyrin repeat domain from Cotesia vestalis polydnavirus (CvBV). Virology 375:374–382

    Article  CAS  PubMed  Google Scholar 

  15. Spencer W, Kwon H, Crepieux P, Leclerc N, Lin R, Hiscott J (1999) Taxol selectively blocks microtubule dependent NF-κB activation by phorbol ester via inhibition of IκBα phosphorylation and degradation. Oncogene 18:495–505

    Article  CAS  PubMed  Google Scholar 

  16. Mikenberg I, Widera D, Kaus A, Kaltschmidt B, Kaltschmidt C (2007) Transcription factor NF-κB is transported to the nucleus via cytoplasmic dynein/dynactin motor complex in hippocampal neurons. PLoS ONE 2:e589

    Article  PubMed  Google Scholar 

  17. Shrum CK, DeFrancisco D, Meffert MK (2009) Stimulated nuclear translocation of NF-κB and shuttling differentially depend on dynein and the dynactin complex. Proc Natl Acad Sci USA 106:2647–2652

    Article  CAS  PubMed  Google Scholar 

  18. Rizki TM, Rizki RM (1994) Parasitoid-induced cellular immune deficiency in Drosophila. Ann N Y Acad Sci 712:178–194

    Article  CAS  PubMed  Google Scholar 

  19. Glatz RV, Asgari S, Schmidt O (2004) Evolution of polydnaviruses as insect immune suppressors. Trends Microbiol 12:545–554

    Article  CAS  PubMed  Google Scholar 

  20. Spradling AC (1993) Developmental genetics of oogenesis. In: Bate M, Martinez-Arias A et al (eds) The development of Drosophila melanogaster. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 1–70

    Google Scholar 

  21. Steinhauer J, Kalderon D (2006) Microtubule polarity and axis formation in the Drosophila oocyte. Dev Dyn 235:1455–1468

    Article  CAS  PubMed  Google Scholar 

  22. Becalska AN, Gavis ER (2009) Lighting up mRNA localization in Drosophila oogenesis. Development 136:2493–2503

    Article  CAS  PubMed  Google Scholar 

  23. Cha BJ, Koppetsch BS, Theurkauf WE (2001) In vivo analysis of Drosophila bicoid mRNA localization reveals a novel microtubule-dependent axis specification pathway. Cell 106:35–46

    Article  CAS  PubMed  Google Scholar 

  24. MacDougall N, Clark A, MacDougall E, Davis I (2003) Drosophila gurken (TGFalpha) mRNA localizes as particles that move within the oocyte in two dynein-dependent steps. Dev Cell 4:307–319

    Article  CAS  PubMed  Google Scholar 

  25. Zimyanin VL, Belaya K, Pecreaux J, Gilchrist MJ, Clark A, Davis I, St Johnston D (2008) In vivo imaging of oskar mRNA transport reveals the mechanism of posterior localization. Cell 134:843–853

    Article  CAS  PubMed  Google Scholar 

  26. Clark I, Giniger E, Ruohola-Baker H, Jan LY, Jan YN (1994) Transient posterior localization of a kinesin fusion protein reflects anteroposterior polarity of the Drosophila oocyte. Curr Biol 4:289–300

    Article  CAS  PubMed  Google Scholar 

  27. Clark IE, Jan LY, Jan YN (1997) Reciprocal localization of Nod and kinesin fusion proteins indicates microtubule polarity in the Drosophila oocyte, epithelium, neuron and muscle. Development 124:461–470

    CAS  PubMed  Google Scholar 

  28. Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:401–415

    CAS  PubMed  Google Scholar 

  29. Rorth P (1998) Gal4 in the Drosophila female germline. Mech Dev 78:113–118

    Article  CAS  PubMed  Google Scholar 

  30. Rubin GM, Spradling AC (1982) Genetic transformation of Drosophila with transposable element vectors. Science 218:348–353

    Article  CAS  PubMed  Google Scholar 

  31. Tautz D, Pfeifle C (1989) A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma 98:81–85

    Article  CAS  PubMed  Google Scholar 

  32. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  PubMed  Google Scholar 

  33. Andrenacci D, Cernilogar FM, Taddel C, Rotoli D, Cavaliere V, Graziani F, Gargiulo G (2001) Specific domains drive VM32E protein distribution and integration in Drosophila eggshell layers. J Cell Sci 114:2819–2829

    CAS  PubMed  Google Scholar 

  34. Prasad M, Jang AC, Starz-Gaiano M, Melani M, Montell DJ (2007) A protocol for culturing Drosophila melanogaster stage 9 egg chambers for live imaging. Nat Protoc 2:2467–2473

    Article  CAS  PubMed  Google Scholar 

  35. Van Doren M, Williamson AL, Lehmann R (1998) Regulation of zygotic gene expression in Drosophila primordial germ cells. Curr Biol 8:243–246

    Article  PubMed  Google Scholar 

  36. Neuman-Silberberg FS, Schüpbach T (1993) The Drosophila dorsoventral patterning gene gurken produces a dorsally localized RNA and encodes a TGF alpha-like protein. Cell 75:165–174

    Article  CAS  PubMed  Google Scholar 

  37. Neuman-Silberberg FS, Schüpbach T (1996) The Drosophila TGF-alpha-like protein Gurken: expression and cellular localization during Drosophila oogenesis. Mech Dev 59:105–113

    Article  CAS  PubMed  Google Scholar 

  38. Schüpbach T (1987) Germ line and soma cooperate during oogenesis to establish the dorsoventral pattern of egg shell and embryo in Drosophila melanogaster. Cell 49:699–707

    Article  PubMed  Google Scholar 

  39. Wasserman JD, Freeman M (1998) An autoregulatory cascade of EGF receptor signaling patterns the Drosophila egg. Cell 95:355–364

    Article  CAS  PubMed  Google Scholar 

  40. Peri F, Bokel C, Roth S (1999) Local Gurken signaling and dynamic MAPK activation during Drosophila oogenesis. Mech Dev 81:75–88

    Article  CAS  PubMed  Google Scholar 

  41. Gonzalez-Reyes A, Elliott H, St Johnston D (1995) Polarization of both major body axes in Drosophila by gurken-torpedo signalling. Nature 375:654–658

    Article  CAS  PubMed  Google Scholar 

  42. Roth S, Neuman-Silberberg FS, Barcelo G, Schüpbach T (1995) Cornichon and the EGF receptor signaling process are necessary for both anterior-posterior and dorsal-ventral pattern formation in Drosophila. Cell 81:967–978

    Article  CAS  PubMed  Google Scholar 

  43. Hawkins NC, Van Buskirk C, Grossniklaus U, Schüpbach T (1997) Post-transcriptional regulation of gurken by encore is required for axis determination in Drosophila. Development 124:4801–4810

    CAS  PubMed  Google Scholar 

  44. Saunders C, Cohen RS (1999) The role of oocyte transcription, the 5′UTR, and translation repression and derepression in Drosophila gurken mRNA and protein localization. Mol Cell 3:43–54

    Article  CAS  PubMed  Google Scholar 

  45. Norvell A, Kelley RL, Wehr K, Schüpbach T (1999) Specific isoforms of squid, a Drosophila hnRNP, perform distinct roles in Gurken localization during oogenesis. Genes Dev 13:864–876

    Article  CAS  PubMed  Google Scholar 

  46. St Johnston D, Nüsslein-Volhard C (1992) The origin of pattern and polarity in the Drosophila embryo. Cell 68:201–219

    Article  CAS  PubMed  Google Scholar 

  47. Berleth T, Burri M, Thoma G, Bopp D, Richstein S, Frigerio G, Noll M, Nüsslein-Volhard C (1988) The role of localization of bicoid RNA in organizing the anterior pattern of the Drosophila embryo. EMBO J 7:1749–1756

    CAS  PubMed  Google Scholar 

  48. Ephrussi A, Dickinson LK, Lehmann R (1991) Oskar organizes the germ plasm and directs localization of the posterior determinant nanos. Cell 66:37–50

    Article  CAS  PubMed  Google Scholar 

  49. McGrail M, Hays TS (1997) The microtubule motor cytoplasmic dynein is required for spindle orientation during germline cell divisions and oocyte differentiation in Drosophila. Development 124:2409–2419

    CAS  PubMed  Google Scholar 

  50. Li M, McGrail M, Serr M, Hays TS (1994) Drosophila cytoplasmic dynein, a microtubule motor that is asymmetrically localized in the oocyte. J Cell Biol 126:1475–1494

    Article  CAS  PubMed  Google Scholar 

  51. Matthies HJ, Baskin RJ, Hawley RS (2001) Orphan kinesin NOD lacks motile properties but does possess a microtubule-stimulated ATPase activity. Mol Biol Cell 12:4000–4012

    CAS  PubMed  Google Scholar 

  52. Cui W, Sproul LR, Gustafson SM, Matthies HJ, Gilbert SP, Hawley RS (2005) Drosophila Nod protein binds preferentially to the plus ends of microtubules and promotes microtubule polymerization in vitro. Mol Biol Cell 16:5400–5409

    Article  CAS  PubMed  Google Scholar 

  53. Shapiro RS, Anderson KV (2006) Drosophila Iκ2, a member of the IκB kinase family, is required for mRNA localization during oogenesis. Development 133:1467–1475

    Article  CAS  PubMed  Google Scholar 

  54. Abdu U, Bar D, Schüpbach T (2006) spn-F encodes a novel protein that affects oocyte patterning and bristle morphology in Drosophila. Development 133:1477–1484

    Article  CAS  PubMed  Google Scholar 

  55. Provost B, Varricchio P, Arana E, Espagne E, Falabella P, Huguet E, La Scaleia R, Cattolico L, Poirié M, Malva C, Olszewski JA, Pennacchio F, Drezen JM (2004) Bracoviruses contain a large multigene family coding for protein tyrosine phosphatases. J Virol 78:13090–13103

    Article  CAS  PubMed  Google Scholar 

  56. Fath-Goodin A, Kroemer JA, Webb BA (2009) The Campoletis sonorensis ichnovirus vankyrin protein P-vank-1 inhibits apoptosis in insect Sf9 cells. Insect Mol Biol 18:497–506

    Article  CAS  PubMed  Google Scholar 

  57. Siegrist SE, Doe CQ (2007) Microtubule-induced cortical cell polarity. Genes Dev 21:483–496

    Article  CAS  PubMed  Google Scholar 

  58. Affolter M, Weijer CJ (2005) Signaling to cytoskeletal dynamics during chemotaxis. Dev Cell 9:19–34

    Article  CAS  PubMed  Google Scholar 

  59. Etienne-Manneville S (2006) In vitro assay of primary astrocyte migration as a tool to study Rho GTPase function in cell polarization. Methods Enzymol 406:565–578

    Article  CAS  PubMed  Google Scholar 

  60. Pan X, Lührmann A, Satoh A, Laskowski-Arce MA, Roy CR (2008) Ankyrin repeat proteins comprise a diverse family of bacterial type IV effectors. Science 320:1651–1654

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

Special thanks go to Carla Malva who inspired this work with her really original and open-minded vision of science. We thank Trudi Schüpbach, Anne Ephrussi, Tom Hays, and Daniel St. Johnston for kindly providing us antibodies and fly strains. We also thank the Bloomington Stock Center for fly stocks and the Developmental Studies Hybridoma Bank for antibodies. We are grateful to Angela Algeri for the careful reading of the manuscript. Our work was supported by grants from the MIUR and University of Bologna (Prin 2006/2008, RFO 2006; 2007). SD had a PhD fellowship from the University of Bologna.

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Authors and Affiliations

  1. Dipartimento Biologia Evoluzionistica Sperimentale, Università di Bologna, Via Selmi 3, Bologna, Italy

    Serena Duchi, Valeria Cavaliere, Luca Fagnocchi & Giuseppe Gargiulo

  2. Istituto di Genetica e Biofisica, CNR, Naples, Italy

    Maria Rosaria Grimaldi, Franco Graziani & Silvia Gigliotti

  3. Dipartimento di Biologia, Difesa e Biotecnologie Agro-Forestali, Università della Basilicata, Potenza, Italy

    Patrizia Falabella

  4. Dipartimento di Entomologia e Zoologia Agraria ‘F. Silvestri’, Università di Napoli ‘Federico II’, Portici (NA), Italy

    Francesco Pennacchio

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  1. Serena Duchi
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  2. Valeria Cavaliere
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  3. Luca Fagnocchi
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Correspondence to Giuseppe Gargiulo.

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18_2010_273_MOESM1_ESM.jpg

Distribution of TnBVANK1 protein. TnBVANK1 protein is localized in the cortical region of the oocyte, as shown by analyzing the oocyte just underneath its surface (A) where the protein shows a granular appearance, which is also detected in the cross section of the same stage 10B egg chamber (B). (JPG 1329 kb)

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Duchi, S., Cavaliere, V., Fagnocchi, L. et al. The impact on microtubule network of a bracovirus IκB-like protein. Cell. Mol. Life Sci. 67, 1699–1712 (2010). https://doi.org/10.1007/s00018-010-0273-2

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