RNA-directed transcriptional gene silencing in plants can be inherited independently of the RNA trigger and requires Met1 for maintenance.
BACKGROUND
The association between DNA methylation and gene silencing has long been recognized; however, signals that initiate de novo methylation are largely unknown. In plants, recognition of RNAs that are inducers of posttranscriptional gene silencing (PTGS) can result in sequence-specific DNA methylation, and the aim of this work was to investigate whether heritable epigenetic changes can occur by this mechanism and if the Met1 methyltransferase is required.


RESULTS
RNA-directed DNA methylation (RdDM) was initiated in 35S-GFP transgenic plants following infection with plant RNA viruses modified to carry portions of either the 35S promoter or the GFP coding region. Targeting of the promoter sequence resulted in both methylation and transcriptional gene silencing (TGS) that was inherited independently of the RNA trigger. Targeting the coding region also resulted in methylation; however, this was not inherited. Expression of Met1 was suppressed in order to investigate its role in initiation and maintenance of RdDM. Initiation of RdDM was found to be Met1-independent, whereas maintenance of methylation and TGS in the subsequent generations in the absence of the RNA trigger was Met1-dependent. Maintenance of methylation associated with systemic PTGS was also found to be Met1-independent.


CONCLUSIONS
RNA-triggered events can lead to heritable changes in gene expression, and it is possible that initiation of other epigenetic phenomena such as trans-silencing and paramutation may have an RNA component.
Jones
L
L
Sainsbury Laboratory, John Innes Centre, Colney Lane, NR4 7UH, Norwich, United Kingdom.
Ratcliff
F
F
Baulcombe
D C
DC
D001483:Base Sequence; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000502:physiology*; D019175:DNA Methylation; D017931:DNA Primers; D020868:Gene Silencing / Q000502:physiology*; D010942:Plant Viruses / Q000502:physiology; D010947:Plants, Toxic; D012328:RNA Viruses / Q000502:physiology; D018749:RNA, Plant / Q000502:physiology*; D014026:Nicotiana / Q000235:genetics* / Q000821:virology; D014158:Transcription, Genetic / Q000502:physiology*
D017931:DNA Primers; D018749:RNA, Plant; D004248:DNA (Cytosine-5-)-Methyltransferases