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A single transgene locus triggers both transcriptional and post-transcriptional silencing through double-stranded RNA production

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Abstract

Silencing of a target locus by an unlinked silencing locus can result from transcription inhibition (transcriptional gene silencing; TGS) or mRNA degradation (post-transcriptional gene silencing; PTGS), owing to the production of double-stranded RNA (dsRNA) corresponding to promoter or transcribed sequences, respectively. The involvement of distinct cellular components in each process suggests that dsRNA-induced TGS and PTGS likely result from the diversification of an ancient common mechanism. However, a strict comparison of TGS and PTGS has been difficult to achieve because it generally relies on the analysis of distinct silencing loci. We describe a single transgene locus that triggers both TGS and PTGS, owing to the production of dsRNA corresponding to promoter and transcribed sequences of different target genes. We describe mutants and epigenetic variants derived from this locus and propose a model for the production of dsRNA. Also, we show that PTGS, but not TGS, is graft-transmissible, which together with the sensitivity of PTGS, but not TGS, to RNA viruses that replicate in the cytoplasm, suggest that the nuclear compartmentalization of TGS is responsible for cell-autonomy. In contrast, we contribute local and systemic trafficking of silencing signals and sensitivity to viruses to the cytoplasmic steps of PTGS and to amplification steps that require high levels of target mRNAs.

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Abbreviations

dsRNA:

Double-stranded RNA

GUS:

ß-Glucuronidase

NIR:

Nitrate reductase

PTGS:

Post-transcriptional gene silencing

siRNA:

Short interfering RNA

ssRNA:

Single stranded RNA

TGS:

Transcriptional gene silencing

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Acknowledgments

We thank Jean-Marie Pollien (INRA, Versailles, France) for all graftings and Allison Mallory (Whitehead Institute, Cambridge, MA, USA) for fruitful discussions. This work was partly funded by the European Union Biotechnology Programm on Gene Silencing (B104-CT96-0253).

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Author notes

  1. Philippe Mourrain

    Present address: INSERM U784 Genetique Moleculaire du Developpement, Ecole Normale Supérieure, 46, rue d’Ulm, 75230, Paris Cedex 05, France

  2. Rik van Blokland

    Present address: Swammerdam Institute for Life Sciences, University of Amsterdam, 1098 SM, Amsterdam, The Netherlands

Authors and Affiliations

  1. Laboratoire de Biologie Cellulaire, Institut Jean-Pierre Bourgin, INRA, 78026, Versailles Cedex, France

    Philippe Mourrain & Hervé Vaucheret

  2. Department of Genetics, FALW-IMC, Vrije Universiteit Amsterdam, De Boelelaan 1087, 1081HV, Amsterdam, The Netherlands

    Rik van Blokland & Jan M. Kooter

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  1. Philippe Mourrain
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  4. Hervé Vaucheret
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Correspondence to Hervé Vaucheret.

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Mourrain, P., van Blokland, R., Kooter, J.M. et al. A single transgene locus triggers both transcriptional and post-transcriptional silencing through double-stranded RNA production. Planta 225, 365–379 (2007). https://doi.org/10.1007/s00425-006-0366-1

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  • DOI: https://doi.org/10.1007/s00425-006-0366-1

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