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Transitive RNA silencing signals induce cytosine methylation of a transgenic but not an endogenous target

Leen Vermeersch (UGent) , Nancy De Winne (UGent) , Jonah Nolf (UGent) , Annick Bleys (UGent) , Aleš Kovařik and Anna Depicker (UGent)
(2013) PLANT JOURNAL. 74(5). p.867-879
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Abstract
Post-transcriptional gene silencing of a primary target gene in plants can coincide with the production of secondary small interfering RNAs (siRNAs) of coding sequences adjacent to the target region and with de novo RNA-directed DNA methylation (RdDM) thereof. Here, we analyzed the susceptibility of transgenic and endogenous targets to RdDM induced by primary and secondary silencing signals. In three different configurations, primary silencing signals were able to direct in trans methylation of chimeric transgenes and the CATALASE2 (CAT2) endogene; however, extensive spreading of methylation occurred only in the transgene, resulting in the methylation of the flanking CAT2 sequence, whereas methylation of the CAT2 endogene was restricted to the target region and the enclosed introns. The secondary silencing signals arising from this transgenic primary target simultaneously silenced a secondary transgene target and the CAT2 endogene, but were only capable of directing RdDM to the transgene. Our data indicate that RdDM is correlated with the in situ generation of secondary siRNAs, occurring in P35S-driven transgenes but not in most endogenes. We conclude that although both endogenes and transgenes are equally sensitive to transitive silencing, differences exist in their susceptibility to undergo secondary RdDM.
Keywords
DIRECTED DNA METHYLATION, DOUBLE-STRANDED RNA, POTATO VIRUS X, Arabidopsis thaliana, post-transcriptional gene silencing, transgene, PLANTS, TRANSCRIPTS, POLYMERASE, SEQUENCES, endogene, GENE, MAINTENANCE, ARABIDOPSIS THALIANA, intron, transitive silencing, RNA-directed DNA methylation

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Chicago
Vermeersch, Leen, Nancy De Winne, Jonah Nolf, Annick Bleys, Aleš Kovařik, and Anna Depicker. 2013. “Transitive RNA Silencing Signals Induce Cytosine Methylation of a Transgenic but Not an Endogenous Target.” Plant Journal 74 (5): 867–879.
APA
Vermeersch, Leen, De Winne, N., Nolf, J., Bleys, A., Kovařik, A., & Depicker, A. (2013). Transitive RNA silencing signals induce cytosine methylation of a transgenic but not an endogenous target. PLANT JOURNAL, 74(5), 867–879.
Vancouver
1.
Vermeersch L, De Winne N, Nolf J, Bleys A, Kovařik A, Depicker A. Transitive RNA silencing signals induce cytosine methylation of a transgenic but not an endogenous target. PLANT JOURNAL. 2013;74(5):867–79.
MLA
Vermeersch, Leen, Nancy De Winne, Jonah Nolf, et al. “Transitive RNA Silencing Signals Induce Cytosine Methylation of a Transgenic but Not an Endogenous Target.” PLANT JOURNAL 74.5 (2013): 867–879. Print.
@article{4179327,
  abstract     = {Post-transcriptional gene silencing of a primary target gene in plants can coincide with the production of secondary small interfering RNAs (siRNAs) of coding sequences adjacent to the target region and with de novo RNA-directed DNA methylation (RdDM) thereof. Here, we analyzed the susceptibility of transgenic and endogenous targets to RdDM induced by primary and secondary silencing signals. In three different configurations, primary silencing signals were able to direct in trans methylation of chimeric transgenes and the CATALASE2 (CAT2) endogene; however, extensive spreading of methylation occurred only in the transgene, resulting in the methylation of the flanking CAT2 sequence, whereas methylation of the CAT2 endogene was restricted to the target region and the enclosed introns. The secondary silencing signals arising from this transgenic primary target simultaneously silenced a secondary transgene target and the CAT2 endogene, but were only capable of directing RdDM to the transgene. Our data indicate that RdDM is correlated with the in situ generation of secondary siRNAs, occurring in P35S-driven transgenes but not in most endogenes. We conclude that although both endogenes and transgenes are equally sensitive to transitive silencing, differences exist in their susceptibility to undergo secondary RdDM.},
  author       = {Vermeersch, Leen and De Winne, Nancy and Nolf, Jonah and Bleys, Annick and Kova\v{r}ik, Ale\v{s} and Depicker, Anna},
  issn         = {0960-7412},
  journal      = {PLANT JOURNAL},
  language     = {eng},
  number       = {5},
  pages        = {867--879},
  title        = {Transitive RNA silencing signals induce cytosine methylation of a transgenic but not an endogenous target},
  url          = {http://dx.doi.org/10.1111/tpj.12172},
  volume       = {74},
  year         = {2013},
}

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