Resistance of RNA-mediated TGS to HC-Pro, a viral suppressor of PTGS, suggests alternative pathways for dsRNA processing.
In plants, double-stranded (ds) RNA that is degraded to small (sm) RNAs that are approximately 23 nucleotides in length can trigger the degradation of homologous RNAs in the cytoplasm (posttranscriptional gene silencing or PTGS) and de novo methylation of homologous DNA in the nucleus [1]. PTGS is similar to quelling in fungi [2] and RNAi in animals [3]. RNA-directed DNA methylation (RdDM) can lead to transcriptional gene silencing (TGS) and the methylation of homologous target promoters if dsRNAs containing promoter sequences are involved [4]. HC-Pro is a plant viral suppressor of PTGS that acts by preventing the accumulation of smRNAs [5, 6] that provide the specificity determinant for homologous RNA degradation [7-10]. Here, we show that HC-Pro does not suppress TGS induced by promoter dsRNA. Moreover, the amount of promoter smRNAs is elevated 5-fold in the presence of HC-Pro, and target promoter methylation is slightly increased without a concomitant rise in the level of promoter dsRNA. The promoter dsRNA, which is not polyadenylated, failed to trigger substantial degradation of polyadenylated, single-stranded promoter RNA. The differential effects of HC-Pro on smRNA accumulation associated with dsRNA-mediated TGS and at least some cases of PTGS suggest that dsRNA processing can occur by alternative pathways, and they support the idea that RdDM is triggered by smRNAs.
Mette
M F
MF
Institute of Molecular Biology, Austrian Academy of Sciences, Billrothstrasse 11, A-5020, Salzburg, Austria.
Matzke
A J
AJ
Matzke
M A
MA
D000594:Amino Acid Oxidoreductases / Q000235:genetics; D000818:Animals; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D019175:DNA Methylation; D020868:Gene Silencing; D008954:Models, Biological; D010944:Plants / Q000235:genetics / Q000378:metabolism / Q000821:virology; D010947:Plants, Toxic; D017800:Potyvirus / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D011401:Promoter Regions, Genetic; D012323:RNA Processing, Post-Transcriptional; D012330:RNA, Double-Stranded / Q000235:genetics / Q000378:metabolism*; D018749:RNA, Plant / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000235:genetics / Q000378:metabolism / Q000821:virology; D014764:Viral Proteins / Q000378:metabolism*
D012330:RNA, Double-Stranded; D018749:RNA, Plant; D014764:Viral Proteins; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase; D003546:Cysteine Endopeptidases; C072775:HC-Pro protein, potyvirus