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Gene silencing in Neurospora crassa requires a protein homologous to RNA-dependent RNA polymerase

Nature volume 399, pages 166–169 (1999)Cite this article

Abstract

In plants and fungi, the introduction of transgenes can lead to post-transcriptional gene silencing1,2. This phenomenon, in which expression of the transgene and of endogenous genes containing sequences homologous to the transgene can be blocked, is involved in virus resistance3,4,5 and genome maintenance6,7. Transgene-induced gene silencing has been termed quelling in Neurospora crassa and co-suppression in plants. Quelling-defective (qde) mutants of N. crassa, in which transgene-induced gene silencing is impaired, have been isolated8. Here we report the cloning of qde-1, the first cellular component of the gene-silencing mechanism to be isolated, which defines a new gene family conserved among different species including plants, animals and fungi. The qde-1 gene product is similar to an RNA-dependent RNA polymerase found in the tomato9. The identification of qde-1 strongly supports models that implicate an RNA-dependent RNA polymerase in the post-transcriptional gene-silencing mechanism. The presence of qde-1 homologues in a variety of species of plants and fungi indicates that a conserved gene-silencing mechanism may exist, which could have evolved to preserve genome integrity and to protect the genome against naturally occurring transposons and viruses.

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Figure 1: Genomic organization of the qde-1 gene.
Figure 2: Sequence alignment of the QDE1 protein with other polypeptides from the SwissProtein sequence database.

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References

  1. Flavell, R. B. Inactivation of gene expression in plants as a consequence of specific sequence duplication. Proc. Natl Acad. Sci. USA 91, 3490–3496 (1994).

    Article  ADS  CAS  Google Scholar 

  2. Depicker, A. & Van Montagu, M. Post-transcriptional gene silencing in plants. Curr. Opin. Cell Biol. 9, 373–382 (1997).

    Article  Google Scholar 

  3. Ratcliff, F., Harrison, B. D. & Baulcombe, D. C. Asimilarity between viral defense and gene silencing in plants. Science 276, 1558–1560 (1997).

    Article  CAS  Google Scholar 

  4. Voinnet, O. & Baulcombe, D. C. Systemic signalling in gene silencing. Nature 389, 553 (1997).

    Article  ADS  CAS  Google Scholar 

  5. Kasschau, K. D. & Carrington, J. C. Acounterdefensive strategy of plant viruses: suppression of posttranscriptional silencing. Cell 95, 461–470 (1998).

    Article  CAS  Google Scholar 

  6. Assaad, F. F., Lee Tucker, K. & Signer, E. R. Epigenetic repeat-induced gene silencing (RIGS) in Arabidopsis. Plant Mol. Biol. 22, 1067–1085 (1993).

    Article  CAS  Google Scholar 

  7. Bingham, P. M. Cosuppression comes to the animals. Cell 90, 385–387 (1997).

    Article  CAS  Google Scholar 

  8. Cogoni, C. & Macino, G. Isolation of quelling-defective (qde) mutants impaired in posttranscriptional transgene-induced gene silencing in Neurospora crassa. Proc. Natl Acad. Sci. USA 94, 10233–10238 (1997).

    Article  ADS  CAS  Google Scholar 

  9. Schiebel, W. et al. Isolation of an RNA-directed RNA polymerase-specific cDNA clone from tomato. Plant Cell 10, 2087–2101 (1998).

    CAS  PubMed  PubMed Central  Google Scholar 

  10. Palauqui, J. C., Elmayan, T., Pollien, J. M. & Vaucheret, H. Systemic acquired silencing: transgene-specific post-transcriptional silencing is transmitted by grafting from silenced stocks to non-silenced scions. EMBO J. 16, 4738–4745 (1997).

    Article  CAS  Google Scholar 

  11. Cogoni, C. et al. Transgene silencing of the al-1 gene in vegetative cells of Neurospora is mediated by a cytoplasmic effector and does not depend on DNA–DNA interactions or DNA methylation. EMBO J. 15, 3153–3163 (1996).

    Article  CAS  Google Scholar 

  12. Stam, M., Mol, J. N. M. & Kooter, J. M. The silence of genes in transgenic plants. Ann. Bot. 79, 3–12 (1997).

    Article  CAS  Google Scholar 

  13. Voinnet, O., Vain, P., Angell, S. & Baulcombe, D. C. Systemic spread of sequence-specific transgene RNA degradation in plants is initiated by localized introduction of ectopic promoterless DNA. Cell 95, 177–187 (1998).

    Article  CAS  Google Scholar 

  14. Metzlaff, M., O'Dell, M., Cluster, P. D. & Flavell, R. B. RNA-mediated RNA degradation and chalcone synthase A silencing in Petunia. Cell 88, 845–854 (1997).

    Article  CAS  Google Scholar 

  15. Wassenegger, M. & Pélissier, T. Amodel for RNA-mediated gene silencing in higher plants. Plant Mol. Biol. 37, 349–362 (1998).

    Article  CAS  Google Scholar 

  16. Baulcombe, D. C. RNA as a target and an initiator of post-transcriptional gene silencing in transgenic plants. Plant Mol. Biol. 32, 79–88 (1996).

    Article  CAS  Google Scholar 

  17. Romano, N. & Macino, G. Quelling: transient inactivation of gene expression in Neurospora crassa by transformation with homologous sequences. Mol. Microbiol. 6, 3343–3353 (1992).

    Article  CAS  Google Scholar 

  18. Cogoni, C. & Macino, G. Conservation of transgene-induced post-transcriptional gene silencing in plants and fungi. Trends Plant Sci. 2, 438–443 (1997).

    Article  Google Scholar 

  19. Schmidhauser, T. J., Lauter, F. R., Russo, V. E. A. & Yanofsky, C. Cloning, sequence, and photoregulation of al-1, a carotenoid biosynthetic gene of Neurospora crassa. Mol. Cell Biol. 10, 5064–5070 (1990).

    Article  CAS  Google Scholar 

  20. Lindbo, J. A., Silva-Rosales, L., Proebsting, W. M. & Dougherty, W. G. Induction of a highly specific antiviral state in transgenic plants: implications for regulation of gene expression and virus resistance. Plant Cell 5, 1749–1759 (1993).

    Article  CAS  Google Scholar 

  21. Montgomery, M. K. & Fire, A. Double-stranded RNA as mediator in sequence-specific genetic silencing and co-suppression. Trends Genet. 14, 255–258 (1998).

    Article  CAS  Google Scholar 

  22. Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature 391, 806–811 (1998).

    Article  ADS  CAS  Google Scholar 

  23. Timmons, L. & Fire, A. Specific interference by ingested dsRNA. Nature 395, 854 (1998).

    Article  ADS  CAS  Google Scholar 

  24. Matzke, M. A., Matzke, A. J. M. & Eggleston, W. B. Paramutation and transgene silencing: a common response to invasive DNA? Trends Plant Sci. 1, 382–388 (1996).

    Article  Google Scholar 

  25. Davis, R. H. & DeSerres, F. J. Genetic and microbiological research techniques for Neurospora crassa. Methods Enzymol. 17, 79–143 (1970).

    Article  Google Scholar 

  26. Vollmer, L. J. & Yanofsky, C. Efficient cloning of genes of Neurospora crassa. Proc. Natl Acad. Sci. USA 83, 4869–4873 (1986).

    Article  ADS  CAS  Google Scholar 

  27. Staben, C. et al. Use of a bacterial Hygromycin B resistance gene as a dominant marker in Neurospora crassa transformation. Fungal Genet. Newslett. 36, 79–81 (1989).

    Google Scholar 

  28. Cabibbo, A., Sporeno, E., Macino, G. & Ballario, P. CBM1, an Neurospora crassa genomic library in pAC3 and its use for walking on chromosome VII right arm. Fungal Genet. Newslett. 38, 68–70 (1991).

    Google Scholar 

  29. Morelli, G., Nelson, M. A., Ballario, P. & Macino, G. Photoregulated carotenoid biosynthetic genes of Neurospora crassa. Methods Enzymol. 214, 412–424 (1993).

    Article  CAS  Google Scholar 

  30. Maniatis, S. T., Fritsch, E. F. & Sambrook, J. Molecular Cloning: A Laboratory Manual (Cold Spring Harbor Laboratory Press, New York, (1982).

    Google Scholar 

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Acknowledgements

We thank G. Azzalin for technical assistance, and G. Coruzzi and A. Pickford for comments on the manuscript. This work was supported in part by grants from the Istituto Pasteur Fondazione Cenci Bolognetti, from the European Union BIOTECH program and from the Ministero dell'Universita' e della Ricerca Scientifica e Tecnologica.

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  1. Dipartimento di Biotecnologie Cellulari ed Ematologia, Sezione di Genetica Molecolare, Universita' di Roma La Sapienza, Viale Regina Elena 324, 00161, Roma, Italy

    Carlo Cogoni & Giuseppe Macino

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  1. Carlo Cogoni
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  2. Giuseppe Macino
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Correspondence to Giuseppe Macino.

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Cogoni, C., Macino, G. Gene silencing in Neurospora crassa requires a protein homologous to RNA-dependent RNA polymerase. Nature 399, 166–169 (1999). https://doi.org/10.1038/20215

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